Introducing:

Human Recombinant Laminin Matrices – Key to Stem Cell BiologyLaminins provide a defined, xeno-free and biorelevant cell culture environment

Biolamina delivers unique new tools for stem cell culture Laminins are the basic foundation of most cells. For the first time, recombinant laminins of human origin are now available, giving cells in culture access to highly-defined, uniquely biorelevant matrices that mimic the natural environment of cell growth. BioLamina’s range of laminin matrices helps you grow both stem cells and differentiated cells, and provides new possibilities to CONTROL, DEFINE and DEVELOP your cell research all the way from lab to clinic.

Reliable data

• Forthefirsttime,arecombinantproteinmatrix combined with a defined cell culture medium gives you full control of cell culture conditions• Acontrolledcellcultureenvironmentletsyourepeat experiments exactly the same way time after time• Fully-definedreagentsletyouchangeculture conditions in a controlled fashion so that their effects can be explored and interpreted with greater confidence

bioRelevant and safe

• Eachofthehumanbody’s15lamininshasaspecial role in cell growth and survival. BioLamina will soon provide all these laminins• Simulateanaturalenvironmentontheculturedishusing specific laminins biorelevant for your particular cell type• Differentlamininssupportspecificcelltypes,which increases safety in clinical trials because the laminin supportingyourdifferentiatedcellsdoesnotsupportEScells

fRom bench to bedside

• RecombinantlamininsprovidedbyBioLaminaare defined and xeno-free • Recombinantproteinscanbemanufactured according to GMP• AllinnovationsperformedwithBioLamina’shuman recombinant laminins can therefore serve as a source for regenerative therapies

FIGURE 1Laminins are extracellular proteins of the basal lamina (basement membrane) located immediately adjacent to cells of most tissue types. The basal lamina is the first extra-cellular matrix produced in the embryo. It forms ultrathin layers beneath epithelial and endothelial cells, or surrounds individual cell types, such as muscle, nerve and fat cells. The basal lamina plays a central role in cell-matrix interactions; it is the foundation for cells to grow on, and it contains molecules, such as laminins, that direct cellular differentiation.Basal lamina closely attaches to cells either:

• Underneathit–asinepithelia and vascular endothelial cells• Aroundit–asinmusclecells, nerve cells and adipocytes

FIGURE 2Laminins are the most abundant proteins in the basement membrane. They are large trimeric proteins that contain an α, β and γ-chain, each of which is found in five, three and three genetic variants respectively. Laminin molecules are named according to their chain composi-tion.Thus,Laminin-511containsα5,β1and γ1chains.

FIGURE 3Laminins bind to cell receptors and connect them with macromolecules of the extracellular matrix. Because they contribute to the foundation of cells, some laminin extracts have been used as a matrix for culturing different cells in vitro, including stem cells. However, certain laminins previously not available to the research community have now been shown to play important, tissue-specific roles in vivo when the cell-matrix interactions involve extensive outside-in signaling. These biologically-active basal lamina proteins influence cell differentia-tion, migration, adhesion, phenotype maintenance and survival, for example.

Figure 2

Laminins are the biorelevant foundation of cell culture

FIGURE 4Sincedifferentcelltypesneeddifferentlaminins in vivo, different cell types also need access to biorelevant laminins to mimic their natural environment in cell culture.Laminin-511,forexample,isalready expressed at the four-cell stage in the developing embryo, and has, together withLaminin-521,beenshowntopromotestem cell proliferation, but not different-iation. In contrast, specialized cells use other laminins for differentiation, adhesion and survival. Thus, epithelial cells need Laminin-332,muscleandnervecellsneedLaminin-211andLaminin-221,andendo-thelial and pancreatic beta-islet cells use Laminin-411combinedwithLaminin-511.Over 7,000 research articles verify the multiple locations and roles of different laminins.

stem cell biology:Embryonicstemcells:LN-511,LN-521

Hematopoieticstemcells:LN-511

immunology:Neurophils:LN-411Thymocytes:LN-511,

LN-332,LN-221

neurobiology:DRGneurons:LN-511Schwanncells:LN-411Motorneurons:LN-211

Toothinnervation:LN-411

Pancreas:Insulin-producing beta cells:

LN-511,LN-411

Kidney:Podocytes:LN-521

muscle: Cardiacmuscle:LN-211/LN-221Skeletalmuscle:LN-211

skin: Keratinocytes:LN-332

vascular system:Vascular endolthelial cells: LN-511,LN-411

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Blood vessel Muscle cellEpithelium

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REFERENCES: Timpletal.Laminin–aglycoproteinfrombasementmembranes.JBiolChem,254:9933-7,1979.Ekblometal.Expressionandbiologicalroleoflaminin-1[laminin-111].MatrixBiol,22:35-47,2003.Aumailleyetal.Asimplifiedlamininnomenclature.MatrixBiol24,326-332,2005.Wondimuetal.Characterizationofcommerciallamininpreparationsfromhumanplacentaincomparisontorecombinantlaminins221,332,511.MatrixBiol,25:(89-93),2006.

Basal Lamina Basal Lamina Basal Lamina

Rapid single-cell suspension expansion of human ES and iPS cellsCulturinghumanembryonicstem(ES)cellsinacontrolled,biorelevantandrepeatablemannerisknowntobe difficult. Moreover, single-cell enzymatic passaging has been possible only by adding apoptosis inhibitors to the cell suspension. BioLamina now presents Laminin-521,anaturalhumanEScellnicheproteinthat allows easy single-cell passaging and large-scale propagationofhumanESandiPScells.

RecombinanthumanLaminin-521isauniqueproteinmatrixthatcreatesabiologi-callyrelevantenvironmenttosupportpluripotentsinglecellgrowthofhumanESandiPScells.Inaddition,handlinghumanpluripotentstemcellcultureshaveneverbeeneasier and more standardized. Combined with an appropriate cell culture medium, thisoffersseveralkeyadvantages:

• Defined,xeno-freeandclonalpropagationofpluripotenthumanESandiPScells for months

• Single-cellpassagingforextremelyeasyandreliablepassagingtechnique– concentrate on your experiments instead of cell culturing

• Nodamagingrhokinase(ROCK)inhibitorneededforcellsurvival

• Enzymaticsingle-cellpassagingforautomationandstandardization

• Biologicalrelevance–Laminin-521isthenaturalnicheforhumanEScells, and is expressed by pluripotent cell during early embryogenesis

• HumanrecombinantLaminin-521canbemanufacturedaccordingtocGMP soyoucantakeyourinnovationfrombenchtobedside

FIGURE 1:SurvivalofdissociatedhEScellswithout ROCK inhibitor on human recombi-nantLaminin-521,mouselaminin-111andMatrigel.CrystalVioletstainingofhEScells24hoursaftersingle-cellsuspensionplatinginmTeSR1mediumwithoutandwithY-27632treatment.

FIGURE 2:HumanEScellculturesexpandrapidly and continuously for months when passaged as single-cells in the absence of ROCK inhibitor. Even better compared to cells grown on Matrigel passaged as clumps.

FIGURE 3: PluripotenthEScellsplatedassingle-cell suspension grow as a monolayer ontopofLaminin-521.Laminin-521mediatesits effect via the α6ß1 integrin, signaling throughthePI3K/Aktpathwayandtheα6ß1integrin isoform is highly expressed in human EScells.Laminin-521hasaveryhighaffinityfor α6ß1and,thus,Laminin-521canconveythehighest dose of biorelevant signal, resulting in increased single-cell survival. The long-term expanded cells homogenously express Oct4(green)andremainkaryotypicallynormal.

REFERENCES: RodinSetal.Poster#3540(Abstract#1901)9thISSCRmeetingJune15-19,2011,Toronto,Canada. RodinS,NiaudetC,HanssonEM,DomogatskayaA,XiaoZ,NilssonAS,HolmF,BakerD,InzunzaJ,RozellB,SunY,BetsholtzC,HovattaO, TryggvasonK.:LN-521permitssurvivalandexpansionofdissociatedhumanpluripotentcellsthroughintegrinα6ß1-mediatedmigrationand PI3K/Aktactivation.Submitted.April2011.

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www.biolamina.com

BioLamina’sbusinessfocusistodevelop,manufactureandmarketproprietary,high-quality, laminin-based substrate solutions for cell growth. Our vision is to set a new golden standard for culturing stem cells and differentiated cell lineages.

Foundedin2008,BioLaminaemanatesfromresearchperformedattheKarolinskaInstitute by the group of Prof. Karl Tryggvason, one of the world’s leading research laboratories in the extracellular matrix field. The company is the only one in the world that manufactures recombinant human laminins.

BioLaminaAB,LöfströmsAllé5A,17266Sundbyberg,Sweden–Tel:+46-8-58885180,Fax+46-8-51989288,Mail:info@biolamina.com

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