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A novel source for Mesenchymal stem cells

Jayanti Tokas1, Deepika Gupta1, Divya Pasrija1, Rubina Begum1, Shalini Jain2and Hariom

Yadav2

1Department of Biotechnology, JMIT, Radaur, Haryana, India

2NIDDK, National Institutes of Health, Bethesda, MD 20892, USA

Corresponding: yadavhariom@gmail.com

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FETAL STEM CELLS

Sources

Specific fetal tissues

Fetal circulation, placenta, Amniotic fluid

Umbilical cord

Potentiality varies from pluripotent to multipotent.

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Fetal stem cells

Hemopoietic stem cells (HSC)

Mesenchymal stem cells (MSC)

First-trimester blood

richer source of HSC and MSC

Greater proliferative capacity

wider differentiation ability

Can be clonally derived.

FETAL STEM CELLS

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AMNIOTIC FLUID

Clear, yellowish liquid, surrounds the fetus during

pregnancy

Continuously inhaled, exhaled and urinated by the

baby.

It is 98% water and electrolytes, proteins, peptides,carbohydrates, lipids and hormones.

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Protects the developing baby against blows to the

mother's abdomen

Allows easier fetal movement

Promotes muscular/skeletal development

Protect the fetus from heat loss.

Plays a significant defensive role

AMNIOTIC FLUID

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Amniotic

fluid

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ROLE IN FETAL DEVELOPMENT

Contains nutritional components such as proteins,

glucose, triglyceride and cholesterol

proteins and peptides possess potent bioactivity

Growth factors in AF are transported throughout

fetal body

Helps cellular movement, organs development,

cellular growth and proliferation.

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AMNIOTIC FLUID STEM CELLS

Potentiality varies from pluripotent to multipotent

Second and third trimester amniotic fluid is major source

Allow scientists to sidestep the controversy over

destroying embryos for research

Can be used for tissue repair and engineering organs

Mesenchymal stem cells quantity is high in amniotic fluid

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HETEROGENEITY OF AMNIOTIC FLUID

Variation in

Shape

Size

Nuclear/ cytoplasmic ratio

Cytoplasmic characteristics

Cell surface properties

Biochemical properties

Some cells express markers of glial and neuronal

stem cells.

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AMNIOTIC FLUID :NEW SOURCE FOR

NEURAL STEM CELLS

Amniotic fluid cells when incubated with neurogenic

medium express

CD133

Nestin

Neurofilament

CNPase

p75

Neurotrophin receptor (the brain-derived neurotrophic factor and

neurotrophin-3)

Human amniotic fluid contains cells that express markers for neuronal

stem and progenitor cells, which harbour the potential to differentiate

into neurogenic cells

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MESENCHYMAL STEM CELLS

Morphology

Small cell body with a few cell processes

Large and round nucleus with a prominent

nucleolus

A small amount of

Golgi apparatus

Rough endoplasmic reticulum Mitochondria

Polyribosomes .

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Detection Surface antigens

Differentiation capacity

Multipotent-Can differentiate into Osteoblasts

Adipocytes

chondrocytes

myocytes

neuron-like cells.

Differentiation capacity decreases with the age ofthe donor and the time in culture

MESENCHYMAL STEM CELLS

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MESENCHYMAL STEM CELLS AND ITS

DIFFERENTIATION CAPACITY

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ISOLATION AND CULTURE

Sterile needle is inserted through the abdominal wall into the

amniotic sac

Small amount of amniotic fluid is withdrawn through needle

Amniotic fluid contains fetal cells which are separated from

amniotic fluid

Cultured

Tests are performed on cultured cells

Chemical analysis

DNA analysis

Chromosomal Analysis

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MESENCHYMAL STEM CELLS : FIRSTTRIMESTER

CD291 CD441

SH21

SH31

SH41

prolyl-4-hydroxylase

Actin

CD452

CD342

CD142

CD682

Vwf2

HLA-DR2

Fibronectin

Laminin

Vimentin

In undifferentiated state, fetal blood, liver and bone marrow

MSCs express

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MSCs differentiate into

Adipocytes

Osteocytes

Chondrocytes.

Provide novel targets for in utero cellular

and gene therapy.

MESENCHYMAL STEM CELLS : FIRSTTRIMESTER

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MSCs exhibit a phenotype and multilineage

differentiation potential similar to postnatal bone

marrow (BM)derived MSCs .

Multipotent (CD29)

Pluripotent (Oct-4)

MESENCHYMAL STEM CELLS : SECONDTRIMESTER

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Express Oct-4.

Samples can be collected in larger amount than the second

trimester, with a much lower risk of uterine contamination.

High renewal capacity and therapeutic applications.

Share same properties and differentiation capabilities as human

embryonic stem cells .

Differ from embryonic stem cells in promising ways :

1. Do not produce teratomas when transplanted into animals.

2. Low antigenicity is an advantage for cell transplantation or cell replacement

therapy.

MESENCHYMAL STEM CELLS : THIRDTRIMESTER

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PRESERVATION

Vitrification

Amniotic stem cell banks

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VITRIFICATION

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AMNIOTIC STEM CELL BANKS

Biocell center, European biotechnology company is

the first firm to harvest and preserve amniotic stem

cells

Biocell Center captures the stem cells from amniotic

fluid if the family requests and pays for the

preservation of the stem cells

Amniotic fluid withdrawn during amniocentesis is

sent to the Biocell Center laboratory. The stem cells

are frozen in liquid nitrogen and preserved in the

company's state-of-the-art cryo-bank.

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H.R. 1892National Amniotic and Placental

Stem Cell Bank of 2007

The bill shows the importance of amniotic stem cells bankwhich would

Establish a National Amniotic and Placental Stem Cell Bank for

the purpose of obtaining, storing, and making available for

research and treatment human stem cells derived from

amniotic fluid or placenta .

Will maintain a collection of at least 100,000 samples of stem

cells in order to ensure genetic diversity

Will obtain stem cells only if informed consent is provided

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REGENERATIVEMEDICINE

As regenerative medicine amniotic fluid stem cells

has many applications .

Cultured MSCs have been transplanted in childrenwith osteogenesis imperfecta (OI), a disease

causing bone fractures and fragility.

Reduced bone fractures and increased bone

density were reported to be found when MSCs were

engrafted into the defective bone

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MYOCARDIAL INFARCTION

MSCs can be engrafted at the damaged

site(s), attenuate pathologic remodeling of

the heart tissue and reduce scar size,

leading to improved post-MI cardiac

function

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ETHICAL ASPECTS

Amniotic fluid solve a lot of problems

It's possible to catch amniotic stem cells without

destroying embryos

They are not necessary to choose an alternative

between donor or autologuous use

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NEXT STEPS TO DO

For the future it is of great importance to obtain more insights into the

spectrum of cells contained in human amniotic fluid.

Using RT-PCR the expression of a wide variety of genes, known as

markers for stem cells, progenitor cells and differentiated cell types,should be analysed.

Amniotic fluid contains Oct-4 positive cells, a major issue for the

future is to investigate the differentiation potential of the Oct-4

positive amniotic fluid cells

It is essential to think about strategies to isolate/enrich Oct-4 positive

cells out of amniotic fluid.

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It is an ocean full of

pearls and we need to

search these pearls only

for our benefit.

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