Autologousregen a morris
Transcript of Autologousregen a morris
A scientific overview and rationale
Development of autologous Adipose Derived Mesenchymal Stem Cell conditioned saline as an injectable cell-free therapy and stem cell banking services
Introduction to the Project
Adult derived mesenchymal stem cells were first isolated
in 1982 and have generated an extensive literature as an
alternative source of stem cells in regenerative medicine.
Mesenchymal stem cells can be isolated from human
bone marrow or from fat tissue of a patient in need of
treatment and thus provide a source of stem cells which
are autologous for the specific patient and so provide a
safer and ethically more acceptable population of stem cells
compared to embryonic stem cells. Despite the 28 years
of extensive research in both animals and humans, there
has been limited progress in translating this research to
therapy. Recently, our traditional view of stem cell therapy
mediating reparative effects in degenerative disease by
direct replacement of damaged cells by healthy cells has
been challenged. Many laboratories including our own have
found that stem cells secrete various factors which rescue
damaged tissue and stimulate regeneration in a variety of
disease models. These data permit an alternative source of
therapy rather than direct stem cell transplantation, namely
the harvesting of such secretory factors in physiological
saline from in-vitro cultured autologous stem cells and using
such saline (conditioned media) as an injectable agent to
ameliorate disease. It is to this approach that this proposal
is addressed. We present examples of our own pre-clinical
animal data for a typical degenerative condition (Dementia),
review the published supporting literature and propose
clinical protocols for assaying such effects in human patients
on a named patient compassionate use basis in a small
private clinic.
Natural Biosciences SA: Company Background.
Natural Biosciences SA. Is a Swiss company, established in
2009, to bring to clinical practice its intellectual property and
expertise in the fields of stem cell derived macromolecules
and regenerative medicine. Our clinical procedures focus
on the use of autologous cell-free injectable lysates and
conditioned media, derived from regulatory authority
compliant adipose derived mesenchymal stem cells, as a
safer (autologous) form
of cell therapy.
A review of the pre-clinical and clinical literature shows an
extensive body of published literature reporting evidence
for cell-free stem cell derived extracts as a safe, minimally
invasive and efficacious therapeutic. Further, many
laboratories including our own, now show such cell free
injections to show the same efficacy as intact stem cell
therapy with less risk to the patient as no intact cells are
transplanted. The approach has been widely used and
stimulates repair and regeneration in a wide range of soft
and hard tissue organs.
The company is focused on two immediate term services,
the facility to offer adults and children the opportunity to
store their own stem cells for future use in the event of
accident or disease. Secondly, to offer clinical therapy using
autologous ADMSC generated cell free therapeutics at our
clinic in Switzerland.
Part 1: Brief overview
Part 1: Overview of therapy and banking services page 2 to 4
Part 2: Scientific rationale for therapy page 5 to 11
Part 3: Resume of Chief Scientist page 12 to 15
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1) Autologous stem cell banking services
There are several well established commercial stem
cell banks offering the opportunity for new parents to store
umbilical cord blood derived stem cells for the child’s use in
later life if the child develops an illness. However, currently
there are no provisions for adult clients to similarly bank
their own stem cells for either future or immediate use in
case of disease or injury. Natural Biosciences offers this
service within the current stem cell regulatory framework to
an international client base, with storage services for intact
stem cell populations suitable for transplantation, and cell
free molecular fractions for immediate use by our consulting
physicians.
Advantages of banking stem cells
• Many acute medical crises e.g. heart attack, stroke
or trauma occur with little or no warning, similarly the
treatment window to optimize repair and regeneration
is brief. To maximize the benefits of regenerative
medicine, pre-stored stem cell product for immediate
use is the only option as isolation, separation,
expansion and therapy after the event would have
significantly less efficacy or none at all.
• Stem cell quality and quantity do deteriorate with age.
Most chronic degenerative conditions e.g. dementias,
Parkinson’s disease, lung diseases, liver and kidney
problems, vascular diseases occur in older individuals
with sub optimal stem cell populations. Logically it is
better to harvest an optimal population when the
client is in good health for future use to exploit the
many repair and regenerative effects of the various
forms of stem cell therapy offered by this company,
and others, such effects relevant to our therapies are
reviewed briefly below.
• Regenerative and anti-ageing medicine has become
one of the fastest growing areas of medical sciences
with new developments announced in the scientific
and popular literature announced daily. Adult stem
cell banking offers the client the opportunity to take
full advantage of current and future developments in
stem cell medicine with their own optimized stem
cells ready for immediate use.
2) Autologous ADMSC generated cell-free therapeutics
Natural Biosciences SA has developed a portfolio of
intellectual property based on the paracrine effects of adult
stem cells. Following an extensive period of pre-clinical
testing in our (UK Government regulated) laboratories, we
are at a phase of development to translate this research to
therapeutic application on a Named Patient Use basis in
Switzerland.
Our approach uses the secretory factors from autologous
stem cells as a therapeutic product. These are generated
from the patients own ADMSCs collected by liposuction,
separated from fat and expanded as clinical grade cells.
These cells are then used to condition sterile physiological
saline. Such conditioned saline may then be condensed
for injection or further fractionated for specific injectables.
One such fractionation is the centrifugal separation of
microvesicles from the conditioned saline. Microvesicles
are becoming appreciated as an important communication
system between damaged tissue and stem cells both
in-vivo and in-vitro. They, like whole stem cell conditioned
saline, can mediate numerous tissue regenerative effects
following non-invasive systemic injection. Such cell free
approaches to therapy, offers many safety advantages over
intact stem cell transplantation minimising the potential risk
of tumour genesis from transplanted cells, immunological
issues or inappropriate stem cell differentiation. The
technique is minimally invasive involving either intravenous
or local injection and is used to support healing following
conventional repair of tissue or as a stand alone therapy to
induce tissue regeneration.
The therapy requires minor aspiration of fat. This is
processed by one of our European compliant laboratories
to produce clinical grade autologous ADMSCs. Following
stringent testing, these cells are used to generate the
autologous stem cell generated cell free injectable products.
These therapies are designed to be used to stimulate
endogenous repair and regeneration mechanisms in the
body and so lend themselves well to support other surgical
and medical interventional programmes.
Brief overview (Cont.)
A further advantage is that the patients’ stem cells may
also be banked cryogenically, and future therapies prepared
immediately from the same sample.
The production of such clinical cell free autologous
products is summarised in figure 1
Fig. 1
The company currently holds a unique market advantage
of owning intellectual property and ‘know how’ for
immediate application in adult stem cell banking
conforming to all regulatory bodies, and is positioned
to offer clients immediate access to autologous cell free
therapy. The cell free autologous injectables will only be
used on a named patient basis in our own clinics and will
not be manufactured or supplied to any third party by
Natural Biosciences SA. We are not seeking to produce a
licensed medicine, only to operate as a private clinic.
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Autologous ADMSC generated cell free therapeutics
Natural Biosciences SA has developed a portfolio of
intellectual property based on the paracrine effects of adult
stem cells. Following an extensive period of pre-clinical
testing in our (UK Government regulated) laboratories, we
are at a phase of development to translate this research
to therapeutic application on a Named Patient Use /
Compassionate trial basis in Switzerland.
Our approach uses the secretory factors from autologous
stem cells as a therapeutic product. These are generated
from the patient’s own ADMSCs collected by liposuction,
separated from fat and expanded as clinical grade cells.
These cells are then used to condition sterile physiological
saline. Such conditioned saline may then be condensed
for injection or further fractionated for specific injectables.
One such fractionation is the centrifugal separation of
microvesicles from the conditioned saline. Microvesicles
are becoming appreciated as an important communication
system between damaged tissue and stem cells both in-vivo
and in-vitro. They, like whole stem cell conditioned saline,
can mediate numerous tissue regenerative effects following
non-invasive systemic injection. Such cell free approaches
to therapy, offers many safety advantages over intact stem
cell transplantation minimising the potential risk of tumour
genesis from transplanted cells, immunological issues or
inappropriate stem cell differentiation.
Advantages of ADMSCs
They are easily and safely obtained from lipoaspirates using
clinically approved commercial kits. Housman (2002) reports
a study by the American Society for Dermatological Surgery
of outpatient cosmetic liposuction performed between 1994
and 2000 showed zero deaths on 66570 procedures and a
serious adverse event rate of 0.68 per 1000 cases (1).
ADMSC are also easy to isolate from lipoaspirates using a
digestion buffer containing 0.1%collagenase (type 1, Sigma)
and 0.25% trypsin (sigma) dissolved in Hank’s buffer
then gravity separation and centrifugation (1500 rpm for
10 minutes) (2). This separation method avoids the use of
magnetic bead or antibody mediated positive or negative
selection of stem cell populations. Purity of cells can also
easily be assessed in ADMSCs by high levels of stem cell
related antigens (CD13, CD29, CD44, CD105, and CD166).
Moreover, the clinical utility is further enhanced compared
to other stem cell types by positive expression of Nanog,
Oct4, Sox-2 and Rex-1, genes normally associated with
embryonic stem cells (2).
The most important feature of ADMSCs is that they may be
obtained in very large, clinically significant, numbers from a
single liposuction procedure without the need to expand the
cells. Fraser et al (2006) report that the frequency of BMSCs
in skeletally mature adults is approximately between 1 in
50,000 and 1 in 100,000 cells in bone marrow aspirates
compared to ADMSCs in lipoaspirates with a frequency of 1
in 100 cells some 500 fold more than found in marrow (3).
Another important advantage of ADMSCs is that they are
easily expanded in tissue culture (2, 3). ADMSCs may be
expanded through at least 25 passages without lose of
stem cell characteristics. Thus, large volumes of ADMSCs
may be obtained.
ADMSCs are also known to have excellent potential to
differentiate in to a wide variety of tissue types an essential
feature for stem cell therapy. Illustrative of this ability,
ADMSCs have been differentiated into Neural tissue (4,5,6),
Cardiac tissue (7,8,9), Skeletal muscle (10,11,12), Cartilage (13,14,15),
Hepatocytes (16), Hematopoietic cells (17), endothelial cells
(18), and Bone (19,20). Thus an expansive literature shows
ADMSCs to be one of the most versatile adult derived stem
cell capable of a wide range of regenerative effects in both
humans and animals.
In conclusion, it is clearly evident from the published
scientific literature and the clinical literature, that autologous
derived ADMSCs offer an attractive source of cells for a
variety of applications in regenerative medicine.
Paracrine effects
Adult derived stem cell transplantation has now been
Part 2: Scientific Rationale for the therapy
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studied and used clinically for well over 26 years; however
it is only in the last five years that scientists and clinicians
have begun to realise that stem cells have more than one
therapeutic benefit. Previously it was thought that injected
stem cells home to tissue damage and replace lost cell
populations, hence the popular rational for transplanting
whole cells. Recently, many authors report impressive
regenerative effects in both animal and human studies which
are mediated by paracrine actions of the stem cells i.e.
secretory molecules which have a rescue/stimulatory effect
on tissue damage in a variety of pathologies.
There are now several seminal papers illustrative of the
clinical potential of using autologous stem cell extracts
or media/saline conditioned by such cells as a safer,
less invasive clinical protocol compared with intact cell
transplantation. Amongst the first papers to report such
effects was the report by Togel et al. (2005) entitled
“Administered mesenchymal stem cells protect against
ischemic acute renal failure through differentiation-
independent mechanisms” (21). These authors conclude
that the profound recovers observed was mediated by
secretory factors produced by the cells and that it was
these growth factors which ameliorated renal damage.
Similar conclusions have now been reported in many
other pathologies including, but not limited to, recovery
of ischemic limb disease by growth factors secreted by
adipose tissue derived stem cells (22). Increasingly the
factors responsible for such effects are being elucidated
in a variety of pathologies including cardiac protection
and functional recovery (23), liver injury repair (24), brain and
spinal tissue damage (25) and many others beyond the
scope of this article; indeed the entire secretion proteome
of mesenchymal stem cells has now been mapped and
reported (26). An ever expanding literature on this topic has
led many groups to investigate direct injection of stem cell
derived lysates to ameliorate tissue damage following spinal
cord injury (27), wound healing (28), tendon repair (29) amongst
many other pathologies. Further, Yeghiazarians et al. (30)
show that cell extract injection into infarcted hearts results
in functional improvement comparable to intact cell therapy.
Similarly, Shabbir et al. report that heart failure therapy can
be mediated by the trophic activities of mesenchymal stem
cells following injection of stem cell conditioned media or
stem cells intra muscularly into the hamstring muscle (31).
They report attenuating heart tissue injury, inhibiting fibrotic
remodelling, and promoting angiogenesis, stimulating
recruitment and proliferation of endogenous tissue stem
cells and reducing inflammatory oxidative stress as some of
the multiple trophic factor effects of MSCs.
Stem cell conditioned media results have been confirmed
by other laboratories around the world in both human and
animal studies including – stem cell conditioned media
induced neuroprotective effects in a model of Huntington’s
disease (32), other laboratories report excellent stem cell
conditioned media effects on injured spinal cords (33), stroke
(34), cardiac damage (35, 36), diabetes (37) and age related
damage (38, 39).
In summary, conditioned media provides a filtered, cell
free, autologous protein and RNA rich saline suitable for
injection to stimulate regeneration. Such conditioned media
is also known to contain microvesicles small membrane
bound structures (20nm – 1µm in diameter) known to be
powerful intercellular communicators with profound ability
to differentiate, proliferate and integrate endogenous and
exogenous stem cells (40,41,42,43,44) and stimulate tissue
repair by a variety of mechanisms (45,46,49). The clinical use of
differentiated or undifferentiated stem cell conditioned media
again provides an attractive, less invasive and cell free
alternative to cell transplantation.
Human use of mesenchymal stem cells
The potential uses of stem cells for tissue repair and
regeneration is beginning to be realised with extensive
clinical trials already completed for adult derived stem
cells and FDA approval for commencement of human
trials for embryo stem cells. An indication of the number of
clinical trials involving stem cells is best illustrated in fig 1.
which shows the number of human stem cell clinical trials
registered with ClinicalTrial.gov. per annum
The 70,000 records are from 165 countries and are
registered federal or privately funded trials. Further, there
are many clinics internationally offering both autologous and non-autologous stem cell therapy
Scientific Rationale for the therapy (Cont.)
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(e.g. www.xcell-centre.com, www.cellmedicine.com,
www.medistem.com, www.emcell.com,
www.tissustemcell.com) these are well established clinical
practices with impeccable safety records using
a variety of stem cell types including umbilical cord stem
cells, bone marrow mesenchymal stem cells or adipose
derived stem cells, by a variety of different injection routes
including intravenous administration and intraspinal injection,
all use intact cells as the injectable material.
Adipose derived mesenchymal stem cells are also currently
used extensively in clinical medicine as intact cell transplants
in breast reconstruction, ligament and tendon repair, bone
defect repair and cosmetic surgery (e.g. www.cytoritx.com,
www.theamarclinic.com, www.facesplus.com,
www.stemnow.com) and also as cell free injectable material
e.g. Sloane Clinic (www.sloaneclinic.com). There are also
extensive publication lists available at www.stemcelldocs.
org reviewing pre-clinical and clinical data.
Similarly, in the peer reviewed literature, mesenchymal stem
cells have been reported to have safe therapeutic potential
following transplantation in humans with osteogenic
imperfecta (78), bone fracture (79), traumatic brain injury (80),
stroke (81), amyotrophic lateral sclerosis (82), graft versus host
disease (83,84), myocardial infarction (85,86). No significant side
effects have been reported.
Sourced from: http://thestemcellblog.com/2009/03/21/
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References cited in document:
1) Houseman, T.S. (2002)
The safety of liposuction: results of a national survey.
Dermatological Surg. 28. 971 – 978.
2) Zhu, Y. et al. (2008)
Adipose-derived stem cells: a better stem cell than BMSC.
Cell Biochemistry and Function. 26. 664 – 675.
3) Fraser, J.K. et al. (2006)
Fat tissue: an unappreciated source of stem cells
for biotechnology.
Trends in Biochemistry. 24(4). 150 – 154.
4) Ashjian, P.H. et al. (2003)
In vitro differentiation of human processed lipoaspirate cells
into early neural progenitors.
Plast. Reconstructive. Surg. 111. 1922 – 1931.
5) Kang, S.K. et al. (2003)
Improvement of neurological deficits by intracerebral
transplantation of human adipose tissue-derived stromal cells
after cerebral ischemia in rats.
Exp. Neurol. 183. 355 – 366.
6) Safford, K.M. et al. (2002)
Neurogenic differentiation of murine and human
adipose-derived stromal cells.
Biochem. Biophys. Research Communications. 294. 371 – 379.
7) Rangappa, S. et al. (2003)
Transformation of adult mesenchymal stem cells isolated from
fatty tissue into cardiomyocytes.
Ann. Thorac. Surg. 75. 775 -779.
8) Gausted, K.G. et al. (2004)
Differentiation of human adipose tissue stem cells using extracts
of rat cardiomyocytes.
Biochem. Biophys. Research Communications. 314. 420 – 427.
9) Planat-Bernard, V. et al (2004)
Spontaneous cardiomyocytes differentiation from adipose tissue
stroma cells.
Circ. Res. 94. 223 – 229.
10) Rodriguez, A.M. et al. (2005)
Transplantation of a multipotent cell population from
human adipose tissue induces dystrophin expression in the
immunocompetent mdx mouse.
J. Exp. Med. 201. 1397 -1405.
11) Mizuno, H. et al. (2002)
Myogenic differentiation by human processed lipoaspirate cells.
Plast. Reconstr. Surg. 109. 199 – 209.
12) Bacou, F. et al. (2004)
Transplantation of adipose tissue derived stromal cells increases
mass and functional capacity of damaged skeletal muscle.
Cell Transplant. 13. 103 – 111.
13) Winter, A. et al. (2003)
Cartilage-like gene expression in differentiated human stem cell
spheroids: a comparison of bone marrow-derived and adipose
tissue-derived stromal cells.
Arthritis Rheum. 48. 418 – 429.
14) Erickson, G.R. et al. (2002)
Chondrogenic potential of adipose tissue-derived stromal cells in
vitro and in vivo.
Biochem. Biophys. Res. Commun. 209. 763-769.
15) Dragoo, J.L. et al. (2003)
Tissue engineered cartilage and bone using stem cells from
human infrapatellar fat pads.
J. Bone Joint Surg. Br. 85. 740-747.
16) Seo, M.J. et al. (2005)
Differentiation of human adipose stromal cells into hepatic lineage
in vitro and in vivo.
Biochem. Biophys. Res. Commun. 328. 258-264.
17) Cousin, B. et al. (2003)
Reconstitution of lethally irradiated mice by cells isolated from
adipose tissue.
Biochem. Biophys. Res. Commun. 301. 1016 – 1022.
18) Miranville, A. et al. (2004)
Improvement of postnatal neovascularisation by human adipose
tissue-derived stem cells.
Circulation. 110. 349 – 355.
19) Zuk, P.A. et al (2001)
Multilineage cells from human adipose tissue: implications foe
cell-based therapies.
Tissue Eng. 7. 211 – 228.
20) Hicok, K.C. et al. (2004)
Human adipose-derived adult stem cells produce osteoid in-vivo.
Tissue Eng. 10. 371 – 380.
21) Togel, F. et al. (2005)
Administred mesenchymal stem cells protect against
ischemic acute renal failure through differentiation
-independent mechanisms.
Am. J. Physiol – Renal Physiol. 289. 31 – 42.
22) Nakagami, H. et al. (2005)
Novel autologous cell therapy in ischemic limb disease through
growth factor secretion by cultured adipose tissue-derived stem cells.
Arterioscler. Thromb. Vasc. Biol. 25. 2542 – 2547.
23) Gneechi, M. et al. (2006)
Evidence supporting paracrine hypothesis for Akt- modified
mesenchymal stem cell-mediated cardiac protection and
functional improvement.
FASEB J. 20. 661 – 669.
24) Lin, N. et al. (2008)
Hedgehog-mediated paracrine interactions between hepatic
stellate cells and marrow derived mesenchymal stem cells.
Biochem. Biophys Res. Comm. 372. 260 – 265.
8
25) Todas. H. et al. (2003)
Stem cell-derived neural stem/progenitor cell supporting factor
for adult neural stem/progenitor cells.
J. Biological Chemistry. 278 (37). 35491 – 35500.
26) Kwan Sze. S. et al. (2007)
Elucidating the secretion proteome of human embryonic
stem cell-derived mesenchymal stem cells.
Molecular & Cellular Proteomics. 6. 1680 – 1689.
27) Kang. S.K. et al. (2007)
Cytoplasmic extracts from adipose tissue stromal cells alleviates
secondary damage by modulating apoptosis and promotes
functional recovery following spinal cord injury.
Brain Pathol. 17. 263 -275.
28) Fu, X. et al. (2007)
Adipose tissue extract enhances skin wound healing.
Wound Repair and Regeneration. 15. 540 – 548.
29) Moon, Y. L. et al. (2008)
Autologous bone marrow plasma injection after arthroscopic
debridement for elbow tendinosis.
Ann. Acad. Med. Singapore. 37. 559 -563.
30) Yeghiazarians, Y. et al. (2009)
Injection of bone marrow cell extract into infracted hearts results
in functional improvement compared to intact cell therapy.
Molecular Therapy. Advanced on line doi:10.1038.mt.2009.85.
31) Shabbir, A. et al. (2009)
Heart failure therapy mediated by trophic activities of bone marrow
mesenchymal stem cells: a non invasive therapeutic regimen.
Am. J. Physiol. Heart. Circ. Physiol. 296. H1888 – H1897.
32) Lim. H.C. et al (2008)
Neuroprotective effect of neural stem cell-conditioned media in in
vitro model of Huntingdon’s disease.
Neuroscience Letters. 435. 175 – 180.
33) Peng, L. et al (2006)
Human neural stem cells promote corticospinal axons regenerating
and synapse reformation in injured spinal cord of rats.
Chinese Medical Journal. 119 (16). 1331 – 1338.
34) Fatar, M. et al. (2008)
Lipoaspirate-derived adult mesenchymal stem cells improve
functional outcome during intracerebral hemorrage by
proliferation of endogenous progenitor cells stem cells in
intracerebral hemorrages.
Neuroscience Letters. 443. 174-178.
35) Doyle. B. et al. (2008)
Progenitor cell therapy in a porcine acute myocardial infarction
model induces cardiac hypertrophy, mediated by paracrine
secretion of cardiotrophic factors including TGFbeta1.
Stem cells and development. 17(5). 941-951.
36) Crisostomo, P.R. et al. (2007)
Stem cell mechanisms and paracrine effects: potential in
cardiac surgery.
Shock. 28(4). 375-383.
37) Moriscot, C. et al. (2005)
Human bone marrow mesenchymal stem cells can express insulin
and key transcription factors of the endocrine pancreas
developmental pathway upon genetic and/or microenvironment
manipulation in vitro.
Stem Cells. 23. 594 – 604.
38) Kim, W.S. et al (2009)
Antiwrinkle effects of adipose-derived stem cells: activation of
dermal fibroblasts by secretory factors.
J. Dermatological Science. 53. 96 – 102.
39) Kim. W.S. et al. (2008)
Evidence supporting antioxidant action of adipose-derived stem
cells: protection of human dermal fibroblasts from oxidative stress.
J. Dermatological Science. 49. 133 – 142.
40) Ratajczak, J. et al. (2006)
Embryonic stem cell-derived microvesicles reprogram
hematopoietic progenitors: evidence for horizontal transfer of
mRNA and protein delivery.
Leukemia. 20. 847-856.
41) Ratajczak, J. et al. (2006b)
Membrane derived microvesicles: important and underappreciated
mediators of cell-to-cell communication.
Leukemia. 20. 1487 – 1495.
42) Yuan, A. et al (2009)
Transfer of microRNAs by embryonic stem cell microvesicles.
PLoS One. 4(3).e4722.
43) Aliotta, J.M. et al (2007)
Alteration of marrow cell gene expression, protein production,
and engraftment into lung by lung-derived microvesicles: a novel
mechanism for phenotype modulation.
Stem Cells. 25(9). 2245 – 2256.
44) Ray.S. (2008)
Patent application Microvesicles. PCT/GB2009/000004
45) Bruno, S. et al. (2009)
Mesenchymal stem cell derived microvesicles protect against
acute tubular injury.
J. Am. Soc. Nephrol. 20. 1053 – 1067.
46) Spees, J.L. et al. (2006)
Mitochondrial transfer between cells can rescue
aerobic respiration.
Proc. Nat. Acad. Sci USA. 103(5). 1283 – 1288.
47) Van Poll, D. et al. (2008)
Mesenchymal stem cell – derived molecules directly modulate
hepatocellular death and regeneration in-vitro and in-vivo.
Hepatology. 47 (5). 1634 – 1643.
48) Hagan, M. et al. (2003) Neuroprotection by human neural
progenitor cells after experimental contusion in rats.
Neuroscience Letters. 351. 149 – 152.
9
49) Hyun Ok Kim1,2, Scong-Mi Choi1, and Han-Soo Kim. (2013)
Mesenchymal Stem Cell-Derived Secretome and Microvesicles as a
Cell-Free Therapeutics for Neurodegenerative Disorders.
Tissue Engineering and Regenerative Medicine. Vol. 10,
No. 3, pp 93-101DOI 10.1007/s13770-013-0010-7
50) Doyle, B. et al (2008)
Progenitor cell therapy in a porcine acute myocardial infarction
model induces cardiac hypertrophy, mediated by paracrine
secretion of cardiotrophic factors including TGFbeta1.
Stem cells and development. 17. 941 – 51.
51) Lanza, C. et al. (2009)
Neuroprotective mesenchymal stem cells are endowed with
a potent antioxidant effect in – vivo.
J. of Neurochemistry. 110. 1674 – 1684.
52) Kyung Kang, S. et al. (2007)
Cytoplasmic extracts from adipose tissue stromal cells alleviates
secondary damage by modulating apoptosis and promotes
functional recovery following spinal cord injury.
Brain Pathology. 17. 263 – 275.
53) Burchfield, J.S. & Dimmeler, S. (2008)
Role of paracrine factors in stem and projenitor cell mediated
cardiac repair and tissue fibrosis.
Fibrosis & Tissue repair. 1:4. 1 – 11.
54) Rehman, J. et al. (2004)
Secretion of angiogenic and antiapoptotic factors by human
adipose stromal cells.
Circulation. 109. 1292 – 1298.
55) Kang, Y et al. (2009)
Proteomic characterization of the conditioned media produced
by the visceral endoderm-like cell line HepG2 and END2: towards
a defined medium for the Osteogenic/chondrogenic differentiation
of embryonic stem cells.
Stem cells and development. 18(1). 77 - 82.
56) Vaca, P. et al. (2006)
Induction of differentiation of embryonic stem cells into
insulin-secreting cells by fetal soluble factors.
Stem Cells. 24. 258 – 265.
57) Kale, V.P. & Limaye. L.S. (1999)
Stimulation of adult human bone marrow by factors secreted
by fetal liver hematopoietic cells: In vitro evaluation using
semisolid clonal assay system,
Stem Cells. 17. 107 – 116.
58) Aliotta, J.M. et al. (2008)
Tissue-specific gene expression of marrow cells co-cultured with
various murine organs. Abstract presented to the American
Society of Hematology 50th meeting. San Francisco.
59) Bentz, K. et al. (2007)
Embryonic stem cells produce neurotrophins in response
to cerebral tissue extract: Cell line-dependent differences.
J. Neuroscience Research. 85. 1057 – 1064.
60) Liu, Y. et al. (2009)
Cell extract from fetal liver promotes hematopoietic differentiation
of human embryonic stem cells.
Cloning and stem cells. 11(1). 51 – 60.
61) Xu, Y.X. (2009) Mesenchymal stem cells treated with rat pancreatic
extract secrete cytokines that improve the glycometabolism of
diabetic rats.
Stem Cell Biology. 41. 1878 – 1884.
62) Di Santo, S. et al. (2009)
Novel cell-free strategy for therapeutic angiogenesis:
in-vitro generated conditioned medium can replace progenitor
cell transplantation.
PLoS ONE 4(5):e5643
63) Planat-Benard, V. et al. (2004)
Spontaneous cardiomyocytes differentiation from adipose tissue
stromal cells.
Circ. Research. 94. 223 – 229
64) Song, Y.H. et al. (2007)
VEGF is critical for spontaneous differentiation of stem cells
into cardiomyocytes.
Biochem. Biophy. Res. Commun. 354. 999 – 1003.
65) Safford, K.M. et al. (2002)
Neurogenic differentiation of murine and human adipose
derived stromal cells.
Biochem. Biophy. Res. Comm. 294. 371 – 379.
66) Safford, K.M. et al. (2004)
Characterization of neuronal/glia differentiation of murine adipose
derived adult stromal cells.
Exp. Neurol. 187. 319 – 328.
67) Ashjian, P.H. et al. (2003)
In-vitro differentiation of human processed lipoaspirate into early
neural progenitors.
Plast. Reconstr. Surg. 111. 1922 – 1931.
68) Zuk, P.A. et al. (2001)
Multilineage cells from human adipose tissue: implications for
cell based therapies.
Tissue Eng. 7. 211 – 228.
69) Mizuno, H. et al. (2002)
Myogenic differentiation by human processed lipoaspirate cells.
Plast. Reconstr. Surg. 109. 199 – 209.
70) Miranville, A. et al (2004)
Improvement of postnatal neovascularisation by human adipose
tissue-derived stem cells.
Circulation. 110. 349 – 355.
71) Planat-Bernard, V. et al. (2004)
Plasticity of human adipose lineage cells toward endothelial cells:
physiological and therapeutic perspectives.
Circulation. 109. 656 – 663.
10
72) Rehman, T. et al. (2004)
Secretion of angiogenic and antiapoptotic factors by human
adipose stromal cells.
Circulation. 109. 1292 – 1298.
73) Halvorsen, Y.D. et al. (2001)
Extracellular matrix mineralization ans osteoblast gene
expression by human adipose tissue derived stromal cells.
Tissue Eng. 7. 729 – 741.
74) Hicok, K.C. et al. (2004)
Human adipose derived stem cells produce osteoids in vivo.
Tissue Eng. 10. 371 – 380.
75) Stewart C A and Morris R G M (1993).
The watermaze. In “Behavioural Neuroscience. A Practical
Approach. Volume I”. Ed A Saghal, IRL Press at Oxford University
press, Oxford, New York, Tokyo, pp107-122.
76) Kuen-Jer, T. et al. (2007)
G-CSF rescues the memory impairment of animal models
of Alzheimer’s disease. J. Expt Med.
On-line www.jem.org/cgi/doi/10.1084/jem20062481.
77) Smith, D.L. et al. (2009)
Reversal of long term dendritic spine alterations in Alzheimer’s
disease models.
Proc. Natl. Acad. Sci. USA. 106. (39) 16877 – 16882.
78) Horwitz, E.M. et al. (2002)
Isolated allogenic bone marrow derived mesenchymal cells
engraft and stimulate growth in children with osteogenesis
imperfecta : implications for cell therapy of bone.
Proc. Natl. Acad.Sci. USA. 99. 8932 – 8937.
79) Bajada, S. Et al. (2007)
Successful treatment of refractory tibial non-union using calcium
sulphate and bone marrow stromal cell implantation.
J. Bone Joint Surg. Br. 89. 1382 – 1386.
80) Zhang, Z.X. et al. (2008)
A combined procedure to deliver autologous mesenchymal
stromal cells to patients with traumatic brain injury.
Cytotherapy. 10. 134 – 139.
81) Bang, O.Y. et al. (2005)
Autologous mesenchymal stem cell transplantation in
stroke patients.
Ann. Neurol. 57. 874 – 882.
82) Ferrero, I. Et al. (2008)
Bone marrow mesenchymal stem cells from healthy donors and
sporadic amyotrphic lateral sclerosis patients.
Cell Transplant. 17. 255 – 266.
83) Ringden, O. et al. (2006)
Mesenchymal stem cells for treatment of therapy resistant graft
versus host disease.
Transplantation. 81. 1390 – 1397.
84) Muller, I. et al. (2008)
Mesenchymal stem cell therapy for degenerative
inflammatory disorders.
Current opinion in Organ Transplantation. 13. 638 – 644.
85) Chen, S.L. et al. (2004)
Effect on left ventricular function of intracoronary transplantation
of autologous bone marrow mesenchymal stem cells in patients
with acute myocardial infarction.
Am. J. Cardiol. 94. 92 – 95.
86) Katritsis, D.G. et al. (2005)
Transcoronary transplantation of autologous mesenchymal stem
cells and endothelial progenitors into infracted human myocardium.
Catheter. Cardiovasc. Interv. 65. 321 – 329.
11
Occupation: Chief Scientist Natural Biosciences SA
Senior Lecturer in Clinical Physiology,
Oxford Brookes University UK
Academic qualifications:
BSc (Hons.) Psychology
Plymouth Polytechnic -1984
PhD Neuroscience
Plymouth Polytechnic-1988
Stephen Ray BSc PhD
Resume
Employment:
2008: Present Chief Scientist Natural Biosciences SA.
2005 – 2008: Chief Scientist Systems Biology
Laboratory Ltd.
2003 – 2005: Chief Scientist Ribostem Ltd.
1993-2003: Senior lecturer in Clinical Physiology School
of Biological & Molecular Sciences, Oxford Brookes
University
Research interests: The biological basis of learning and memory, neural
tissue transplantation, stem cell transplantation, cell and
cell extract based therapy, physiological measurement
of stress.
12
Selected research publications
Henry Collins-Hooper, Roberta Sartori, Raymond
Macharia, Komtip Visanuvimol, Keith Foster, Antonios
Matsakas, Hannah Flasskamp, Steve Ray, Philip
R Dash, Marco Sandri, and Ketan Patel (2013).
Propeptide-Mediated Inhibition of Myostatin Increases
Muscle Mass Through Inhibiting Proteolytic Pathways in
Aged Mice, Journal of Gerontology A Biol Sci Med Sci.
1 - 11.
Henry Collins-Hooper, Graham Luke, Mark Cranfield,
William R. Otto, Steve Ray, and Ketan Patel. (2011)
Efficient myogenic reprogramming of adult white fat
stem cells and bone marrow stem cells by freshly
isolated skeletal muscle fibers. Translational Research.
Vol. 158 (6) 334 - 343
Walthall H, and Ray S. (2002)
Do perioperative variables have an effect on extubation
timing following coronary artery bypass grafts? Heart
and Lung. Vol. 31.
Walthall H, Robson D, and Ray, S. (2001)
Do any preoperative variables affect extubation time
after coronary artery bypass grafts? Abstract selected
by Dannemiller Memorial Educational Foundation
for AnesthesiaFile.
Walthall, H., Robson, D., & Ray, S. (2001)
Do any preoperative variables affect extubation time
after coronary artery bypass graft surgery?
Heart & Lung. Vol. 30. (3). 216-224.
Walthall H, Robson D, and Ray S. (2001)
Does extubation cause haemodynamic instability
in patients following coronary artery bypass grafts?
Journal of Intensive and Critical Care Nursing.
Vol. 17. 286 – 293.
Ray, S. & Bagnall, L. (2001)
Gender differences in rat learning assessed on
spatial navigation, taste aversion and visual
discrimination tasks.
Animal Technology. Vol. 52. (3). 219 – 226.
13
Martin, M. & Ray, S. (2001)
A comparison of learning ability between water or food
deprived and non deprived rats on
visual discrimination and conditioned taste
aversion paradigms.
Animal Technology. Vol. 52. (3) 211- 217.
Butcher, O. & Ray, S. (2000)
Olfactory learning in the neonate rat: a comparison
of aversive and appetitive conditioning.
Animal Technology. Vol. 51. (3). 151 – 160.
Butcher, O. & Ray, S. (2000)
Development of a suitable in-vivo technique to
explore the effects of Brain derived extracts on neonate
rat maturation.
Animal Technology. Vol. 51 (2). 91- 100.
Ferneyhough, B.M. & Ray, S. (2000)
Long term captivity and its effects on olfactory learning
in the Honeybee Apis mellifera.
Journal of Apicultural Research.
Ferneyhough, B.M. & Ray, S. (1999)
The Honey bee as a laboratory animal in the study of
the biological basis of behaviour.
Animal Technology. Vol. 50. (3). 145 – 154.
Bagnall, L. & Ray, S. (1999)
Rat strain differences on performance in the
Morris water maze.
Animal Technology. Vol. 50 (2). 69 - 77.
Ray, S. & Ferneyhough, B.M. (1999)
Behavioural development and olfactory learning in the
Honeybee (Apis mellifera).
Developmental Psychobiology. Vol. 34. 21 - 27.
Ray, S. (1999)
Survival of Olfactory memory through metamorphosis in
the fly Musca domestica.
Neuroscience letters. Vol. 259. 37 - 40.
Ray, S. (1998)
An alternative to water deprivation techniques in animal
learning studies.
Animal Technology. Vol. 49 (3). 113 – 120.
Ray, S. (1998)
Learning in the land snail Helix aspersa.
Animal Technology. Vol. 49 (3). 135 – 143.
Schofield, L., Ferguson, J.C. & Ray, S. (1997)
Physiological measurement of the response to and
recovery from stress during a selection procedure.
Proceedings of the British Psychological Society.
Vol. 5 (2). 114.
Ray, S. & Ferneyhough, B.M. (1997)
Seasonal variation of proboscis extension reflex
conditioning in the honey bee (Apis mellifera).
Journal of Apicultural Research. Vol. 36 (2). 108 – 110.
Ray, S. & Ferneyhough, B.M. (1997)
The effects of age on olfactory learning in the honey bee
Apis mellifera.
Neuroreport. Vol 8. 789 – 793.
Schofield, L., Ferguson, J.C. & Ray, S. (1997)
Physiological measurement of the response to and
recovery from stress during a selection procedure.
Proceedings of the British Psychological Society. Vol.
5 (2). 114.
Ray, S. (1993)
Recovery from stress measured by Salivary IgA. British
Psychophysiology Society Newsletter. Vol 20. 33.
Resume (Cont.)
14
Selected conference papers
Keeton, S.J., Ray,S. & Dash, P.R. (2013) Plasticity of cell migration st transition zones between different dimensions and substrates. Invadosomes Conference Nijmegen.
Ray, S. (2012) Microvesicles and Tissue repair. NeuroRehab*
Ray, S. (2011) Therapeutic cell banking applications of adipose derived Mesenchymal stem cells in anti-aging or regenerative medicine. Institute of South African Plastic Surgeons National Meeting. *
Ray, S. (2011) Stem cells and neurological repair. NeuroRehab Conf. Berkshire.*
Ray, S. (2009) Paracrine stem cell mediated repair of tissue Damage. Muscular Dystrophy UK.
Ray, S. (2007) Stem cell Repair in Neuromuscular Diseases. Ataxia Society.
Ray, S. (2007) Stem Cell Therapy in Degenerative Diseases. World Anti-aging Medicine Conference . Athens.
Ray, S. (2004) Cell Therapy: promises, promises, promises! Multiple Sclerosis Society Annual Conference.
Ray, S. (2004) Potential applications of cell therapy for neurodegenerative diseases. MS Therapy Centre Annual Lecture. Berkshire.*
Ray, S. (2003) Stem cells and the future of regenerative medicine. Multiple Sclerosis Society. Henley on Thames.
Ray, S. (2002) RNA, are memories made of this? Michaelmas Lecture. Oxford Graduate Society. Oxford
Ray, S. (2002) Potential applications of cell therapy for neurodegenerative diseases. Motor Neurone Disease Association. Nottingham.
Ray. S. (2002) Stem cell therapy in neurodegenerative diseases: an update. Motor Neurone Disease Association Annual Conference, Birmingham.*
Ray, S. (2002) Generation of glia from Bone Marrow stem cells in – vitro. Applications in the treatment of Multiple Sclerosis. Multiple Sclerosis Society Regional Research Conference.
Ray, S. (2001) Macromolecular theories of memory. Institute of Biologists. Reading.
Ray, S. (2001) Stem cell therapy: the use of Bone Marrow Stromal stem cells for global cell replacement in neurodegenerative disease. Multiple Sclerosis Society Research Conference. Manchester.*
Ray, S. (2001) The potential application of stem cell therapy in Motor Neurone Disease. Motor Neurone Disease Association Annual Conference. Birmingham.*
Ray, S. & Freeman-May, A. (2001) Stress profiling of emergency medical staff: A physiological perspective. Ambulance Research & Development, University of Hertford.
Ferneyhough, B.M. & Ray. S. (2001) The effects of behavioural development on olfactory learning and memory in the honeybee (Apis mellifera). Experimental Analysis of Behaviour Conference, University of London.
Martin, M. & Ray. S. (2001) Taste aversion learning in the rat in the absence of food/ water deprivation. Institute of Animal Technology Congress. Jersey.
Ray, S. (2000) The Anatomy of the creative human brain. YPSL European Quality Conference. Thames Valley University.
Ray, S. (1998) Physiological profiling of the stress response in occupational medicine. IPD Conference. Harrogate.*
15
Bagnall, L. & Ray, S. (1998) Rat strain differences in spatial learning assessed on the Morris water maze. Institute of Animal Technology Congress. Jersey.
Ray, S., Howells, K.F., Abbas, S., Butcher, O.L. & Bagnall, L. (1998) The effects of brain derived protein fractions on recipient brain anatomy and behaviour following systemic injection in the rat. Joint meeting of the European Neuropeptide Club and the Summer Neuropeptide Conference, University of Gent, Belgium.
Ray, S. & Ferneyhough, B. (1998) The involvement of neuropeptides in the survival of memory through insect metamorphosis. Joint meeting of the European Neuropeptide Club and the Summer Neuropeptide Conference, University of Gent, Belgium.
Ferneyhough, B. & Ray, S. (1998) Neuropeptides and the ontogeny of learning in the honeybee. Joint meeting of the European Neuropeptide Club and the Summer Neuropeptide Conference, University of Gent, Belgium.
Butcher, O.L. & Ray, S. (1998) The effects of exogenous neuropeptides on the development of motor co-ordination in the neonate rat. Joint meeting of the European Neuropeptide Club and the Summer Neuropeptide Conference, University of Gent, Belgium.
Abbas, S. & Ray, S. (1998) An in-vitro assay for neuropeptide induced plasticity in neuronal and glial transplantation. Joint meeting of the European Neuropeptide Club and the Summer Neuropeptide Conference, University of Gent, Belgium.
Schofield, L.A., Ferguson, J.C. & Ray, S. (1997) Physiological measurement of the response to and recovery from stress during selection procedures. British Psychological Society Occupational Psychology Conference.
Ray, S. (1997) The Physiology of Stress. IPD Annual Conference. Harrogate.
Ray, S., & Blythe J. (1997) Survival of memory through metamorphosis, Experimental Analysis of Behaviour Conference, University of London.
Ray, S. (1997) Neural transplantation and learned time signals, Experimental Analysis of Behaviour Conference, University of London.
Patents
1. BBSRC industrial Studentship Professor Ketan Patel,
Dr Steve Ray Stem cell generated paracrine Therapy
2. BBSRC industrial Studentship Professor Ketan Patel
(University of Reading), Dr Dionne Tannetta
(University of Oxford & Dr Steve Ray (Natural
Biosciences Ltd) Characterising the role of
microvesicles as anti-aging reagents through their
ability to clear protein aggregates
3. BBSRC industrial Studentship Dr Phil Dash &
Dr Steve Ray Microvesicle shedding in cell migration
4. Myostis Support Group Research Grant
Dr Steve Ray, Stem cell microvesicles and
muscle repair
5. BBSRC Studentship Professor Ketan Patel,
Dr Steve Ray, Dr Henry Collins-Hooper, The role of
RNA in stem cell differentiation abd tissue
regeneration
Ray, S. (2003) GB0316089.2 Method of altering cell properties by Administering RNA.
Ray, S. (2004) PCT.GB2004/00298. Method of altering cell properties by Administering RNA.
Ray, S. & Fischer, M. (2006) WO2006077409. Method of genotypic modification by administration of RNA.
Ray, S. (2008) GB0804932.2 Microvesicles
Ray, S. (2009) WO/2009/087361 Microvesicles
* Key Note Speaker
Natural Biosciences SA155A Seestrasse,Kilchberg 8802ZurichSwitzerlandT +41 (0) 44 716 4828 natural-biosciences.com