Science Room

Clonal Cell Culture Applications

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LAMININ-521


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PROTOCOL

Protocol for defined and xeno-free clonal culture of hESC and iPSC on laminin-521 here


VIEW ON DEMAND

Defined and xeno-free cloning and long-term expansion of hESC and iPSC on laminin-521

Watch webinar on defined and xeno-free clonal culture and expansion of hESC and iPSC on laminin-521 here


APPLICATION NOTE

Get the hPSC culture application note here


CUSTOMER TESTIMONIALS

"BioLamina's human recombinant laminin-521 is outstanding in supporting human embryonic stem cell growth. In addition, it makes hES cell handling easy because of single-cell passaging with trypsin."

                                       Prof. Outi Hovatta, Karolinska Institute, Sweden 

         
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ARTICLES

Find a selection of hPSC clonal culture articles here


MORE INFORMATION

Defined and reliable expansion of hPSC on laminin-521

EFFICIENT CLONAL CELL CULTURE ON LAMININ SUBSTRATES

Robust and reliable clonal expansion of human ES and iPS cell expansion on defined and xeno-free human recombinant laminin-521 cell culture reagentsLaminin-521 successfully replicates the genuine human stem cell niche in vitro

In the developing embryo, the alpha-5 laminins (laminin-511 and laminin-521) are the first extracellular matrix proteins to be expressed around the stem cells in the inner cell mass and support survival and self-renewal of the pluripotent stem cells. The alpha-5 laminins are also produced endogenously by pluripotent stem cells cultured in vitro and are critical autocrine and paracrine factors that regulates their survival and self-renewal. 

Human recombinant laminin-521 is a chemically defined and xeno-free system for robust, long-term expansion of human pluripotent stem cells. It successfully replicates the genuine human stem cell niche in vitro and thereby provides a highly reliable and reproducible culture system to create sufficient populations of undifferentiated cells that, in turn, can be terminally differentiated. 


Efficient generation of clinically compliant human ES cell lines on laminin-521

In a publication by Rodin et al., the authors show that laminin-521 supports efficient generation and large-scale expansion of clinically compliant human ES cell (hESC) lines. The biorelevant laminin-521 clonal stem cell culture protocol vastly improves survival of completely dissociated human ES and iPS cells, even at clonal density. The cells quickly adhere to the laminin-521 matrix and start to divide with high survival. Laminin-521 matrix in combination with E-cadherin, even allows clinically compliant hESC line derivation from single blastomere cells without a need to destroy the embryo.

High-efficiency iPSC reprogramming and expansion on laminin-521

Laminin-521 is an optimal defined, xeno- and feeder-free matrix for the clinically compliant reprogramming and expansion of human iPS cells. Reprogramming on laminin-521 has successfully been performed by mRNA and Sendai-virus methods. Lu et al. show that laminin-521 achieves high-efficiency reprogramming and rapid, long-term self-renewal of iPSC with normal karyotype and pluripotency. Moreover, laminin-521 supported direct hiPSC lineage-specific differentiation. 

Tano and colleagues show a novel approach where they use laminin-521 for direct and sensitive detection of trace amounts of residual undifferentiated hPSCs for cell therapy products. The highly efficient culture system detected colony forming hPSCs spiked into primary human MSCs or neurons at a ratio as low as 0.001%–0.01%.

A publication by Matsumura and co-workers show how laminin-521 increase survival of single-cell hiPSC clones. The authors describe increased survival and propagation of single hiPSC clones on laminin-521 in both culture and on a new microfluidic device by conditioning the medium.


Robust, long-term expansion of human ES and iPS cells on laminin-521

A detailed step-by-step protocol for secure transfer from feeders, true clone generation and expansion of hPSCs and freezing and thawing as single cells on LN-521 has been published by Rodin et al. There are also critical steps and reagents included for easier handling of more difficult lines and a useful trouble shooting guide.

Once the cell line is established, LN-521 supports robust, long-term, single-cell expansion of human pluripotent stem cells. Pluripotent stem cells grow as a monolayer on LN-521 providing you with a homogenous, pluripotent cell culture without need to dispose of unwanted differentiated cells. Cells amplify faster compared to other matrices, with about 10-fold expansion in 4 days. Due to the authentic biorelevant signals provided by the culture matrice, laminin-521 is a flexible culture system. It works with most commercial medias, the cells can be passaged in very low cell densities and can be cultured to high confluence without phenotypic alterations or genetic alterationsand. Moreover, hPSCs can routinely be cultured as single cells on laminin-521 without need to add apoptosis inhibitors, such as ROCKi. Since laminin efficiently buffers endogenously produced growth factors LN-521 also performs well together with growth-factor reduced medias. The biorelevant support provided by the laminin-521 matrix also have a protective effect against DNA damage caused by suboptimal culture conditions, such as high-density culture and pH changes. This allows for weekend-free feeding, drastically reducing work load, time and cost for growing your cells.

LAMININ KEY ADVANTAGES

  • Defined and xeno-free cell culture matrix for clinical compliance

  • A biologically relevant culture environment for human ES and iPS cells 

  • No lot-to-lot variability for standardized experiments with less variation

  • High clonal efficiency for hESC derivation, iPSC reprogramming and genome editing

  • Rapid cell expansion for fast scale-up

  • Homogenous and genetically stable hPSC cultures without need to remove differentiated parts 

  • Triggers the authentic cell signaling pathways which generates hPSCs with a more uniform gene expression profile

  • Laminin-521 primes the hPSCs and gives more efficient differentiation and enhances cell maturation, polarization and organization

  • Flexible culture system and easy to control: compatible with any medium, supports weekend-free feeding, single-cells passage without ROCKi, low density seeding to high confluence culture

  • Scientifically validated in high-impact journals

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  • Expansion of human PSC

  • Clonal cell culture applications

  • Eye cells

  • Cardiac cells

  • Neural cells

  • Skeletal muscle cells

  • Kidney cells

  • Hepatic cells

  • Cancer cells

  • Lung cells

  • Intestinal cells

  • Pancreatic cells

  • Hematopoietic stem and progenitor cells

  • Mesenchymal stem cells

  • Animal stem cells

  • Endothelial cells

  • Normal and cancerous mammary cells

  • Epithelial cells

Laminin-521

Laminin-521 human stem cell matrix makes pluripotent stem cell culture easy in a defined, xeno-free and biologically relevant cell culture system.

  • x
  • Neural cells

  • Hepatic cells

  • Lung cells

  • Intestinal cells

  • Normal and cancerous mammary cells

  • Expansion of human PSC

  • Clonal cell culture applications

  • Eye cells

  • Cardiac cells

  • Skeletal muscle cells

  • Kidney cells

  • Cancer cells

  • Pancreatic cells

  • Hematopoietic stem and progenitor cells

  • Mesenchymal stem cells

  • Animal stem cells

  • Endothelial cells

  • Epithelial cells

Laminin-111

Laminin-111 is important during early epithelial development and differentiation of the epiblast, and is commonly used as a general attachment protein in cell culture.

  • x
  • Neural cells

  • Eye cells

  • Endothelial cells

  • Animal stem cells

  • Pancreatic cells

  • Epithelial cells

  • Cardiac cells

  • Cancer cells

  • Hepatic cells

  • Intestinal cells

  • Kidney cells

  • Lung cells

  • Skeletal muscle cells

  • Expansion of human PSC

  • Clonal cell culture applications

  • Hematopoietic stem and progenitor cells

  • Mesenchymal stem cells

  • Normal and cancerous mammary cells

LAMscreen

LAMscreen is a kit with all our currently available laminin isoforms so that you can easily screen the best laminins for different cell types.