Science Room

hPSC Derivation, Maintenance and Cryopreservation

PRODUCT RECOMMENDATIONS

LAMININ-521
 
Buy human recombinant laminin-521 cell culture matrix here                                             Go to the BioLamina shop

 

PROTOCOLS

 

VIEW ON DEMAND

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

Watch webinar on long-term culture of hESC and iPSC on laminin-521 cell culture substrates under defined and xeno-free conditions here

 

hPSC cultured on laminin-521 show decreased proneness to acquire DNA damage during suboptimal culture conditions

HPSC CULTURED ON LAMININ-521 SHOW DECREASED PRONENESS TO ACQUIRE DNA DAMAGE DURING SUBOPTIMAL CULTURE CONDITIONS 



APPLICATION NOTES

 

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 


"We have discovered L521 is far superior to all other alternative ECMs we have tried for hPSC culture, and have switched all our hiPSC and hESC lines to L521 for maintenance. As a result our hPSC lines have become more stable, yields increased and survival improved dramatically on passage. We can now perform more experiments with each flask, and do so with greater confidence of high quality results due to the morphological uniformity of pluripotent cells grown on L521."

                                    Dr. Jonathan Niclis, Univ. of Copenhagen, Denmark

         
MORE TESTIMONIALS HERE

 


ARTICLES

hPSC CULTURE ARTICLES

 

MORE INFORMATION

              Cost-effective hPSC culture on laminin-521

 

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

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

  • Rapid cell expansion for fast scale-up

  • Clonal culture

  • Laminin-521 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

     

Robust and reliable 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-1 and alpha-5 laminins are the first extracellular matrix proteins to be expressed. Laminin-111 is mostly expressed in extra-embryonic basement membranes and stimulates cells to commit to differentiation. Contrary, laminin-521 and laminin-511 are expressed around the stem cells in the inner cell mass and support survival and self-renewal of the pluripotent stem cells (Domogatskaya, 2012; Rodin, 2014a). The alpha-5 laminins are also produced endogenously by human pluripotent stem cells (hPSC) cultured in vitro and are critical autocrine and paracrine factors that regulates their survival and self-renewal (Laperle, 2015). 

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 (Rodin, 2014a). 

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

The natural cell-matrix interaction triggers authentic biorelevant signals and gives homogenous and genetically stable PSC cultures with a more uniformed gene expression. The cells migrate quickly on laminin-521, which correlates with the high survival of the cells. Cells amplify faster compared to other matrices, with about 10-fold expansion in 4 days (Rodin, 2014a). Laminin-521 also support high post-thawed survival of cryopreserved pluripotent stem cells (Miyazaki, 2013). Due to faster growth rate and higher cell yield, the laminin-521 culture system is more cost-effective compared to other feeder-free matrices.

Human ES and iPS cells grow as a monolayer on laminin-521, providing you with a homogenous, high-quality cell culture (Rodin, 2014a; . Compared to conventional colony passaging, hPSCs can be cultured on laminin-521 to high confluency without risk of spontaneous differentiation and there is no need to clear the culture from unwanted, differentiated cells (Rodin, 2014a; Rodin, 2014b).  

Laminin-521 regulates signaling pathwyas that promotes hPSC survival and self-renewal 

Laminin-521 has strong interaction to the α6β1 integrin which induces the PI3K/Akt signaling pathway. In pluripotent stem cells, this interaction promotes survival and efficient long-term self-renewal (Rodin, 2014a; Rodin, 2014b). Activation of integrin α6β1 by alpha-5 laminins have also show to regulate focal adhesion kinase (FAK) signaling in hPSCs and disruption of this pathway results in hPSC differentiation (Villa-Diaz, 2016). Laminin-511 binds the same integrin but the α6β1 integrin mediating effects of laminin-521 is much stronger than that of laminin-511 which results in more robust hPSC expansion on laminin-521 (Rodin, 2014a). 

A flexible culture system that is easy to control

Due to the authentic biorelevant signals provided by the culture matrice, laminin-521 is a flexible culture system. Laminin-521 works with most commercial medias and hPSCs can routinely be cultured as single cells without need to add apoptosis inhibitors, such as ROCKi. Since laminin efficiently buffers endogenously produced growth factors laminin-521 also performs well together with growth-factor reduced medias (Rodin, 2014a). 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 (Jacobs, 2016). This allows for weekend-free feeding, drastically reducing work load, time and cost for growing your cells. 

Laminin-521 supports clonal culture

With the support from the laminin-521 matrix, cells can be passaged in very low cell densities and can be cultured to high confluence without phenotypic alterations (Rodin, 2014a; Rodin, 2014b). Laminin-521 also supports efficient clonal culture and is an excellent substrate for iPSC reprogramming (Uhlin, 2017; Tano, 2014; Lu, 2014). Clinically compliant human embryonic stem cell lines can even be derived from a single blastomere and there is no longer any need to destroy the embryo (Hovatta, 2014; Damdimopoulou, 2015). Importantly, culture on laminin-521 primes the hPSCs and gives more efficient differentiation and enhances cell maturation, polarization and organization (Cameron, 2015; Penton, 2016).

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. The authors also provide data around critical steps and reagents included for easier handling of more difficult lines and a useful trouble shooting guide (Rodin, 2014b).


RECOMMENDED PUBLICATIONS


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More articles HERE

  • x
  • 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

  • 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

  • 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

  • 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.