Biorelevant culture of human ES and iPS cells on Biolaminin substrates
Biolaminin 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 regulate their survival and self-renewal (Laperle, 2015).
Human recombinant laminin 521, Biolaminin 521 LN (LN521) is a chemically defined and animal origin-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).
Biolaminin 521 homogenizes and stabilizes pluripotent gene expression profiles between human ES cell lines
In an article by Albalushi et al., the authors tested the effect of the LN521 substrate on cultured hES cells. Five different human embryonic stem (hES) cell lines, originally derived on human foreskin fibroblasts (hFFs), were cultured on hFF, Matrigel, or laminin-521. Culture on LN521 significantly reduced the variation of pluripotency marker expression compared to culture on hFFs. Moreover, hES cells cultured on LN521 were more homogenous, attached better, and grew faster compared to hES cells cultured on Matrigel (Albalushi, 2017). The results show a positive effect of LN521 in homogenizing and stabilizing pluripotent gene expression profiles between hES cell lines and provide the first step towards generating more controllable, robust, and clinically safe culture conditions.
Robust, long-term expansion of high-quality human ES and iPS cells on LN521
The natural cell-matrix interaction triggers authentic biorelevant signals and gives homogenous and genetically stable PSC cultures with more uniformed gene expression. The cells migrate quickly on LN521, 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). LN521 also supports high post-thawed survival of cryopreserved pluripotent stem cells (Miyazaki, 2013). Due to faster growth rate and higher cell yield, the LN521 culture system is more cost-effective compared to other feeder-free matrices.
Human ES and iPS cells grow as a monolayer on LN521, providing you with homogenous, high-quality cell culture (Rodin, 2014a). Compared to conventional colony passaging, hPSCs can be cultured on LN521 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).
LN521 regulates signaling pathways that promote hPSC survival and self-renewal
LN521 has a strong interaction with the α6β1 integrin, a key mediator of signaling pathways that regulate, survival, proliferation, migration, and differentiation. LN521 binding to the α6β1 which induces the PI3K/Akt signaling pathway which in pluripotent stem cells promotes survival and efficient long-term self-renewal (Rodin, 2014a; Rodin, 2014b). Activation of integrin α6β1 by alpha-5 laminins has also shown 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, LN521 is a flexible culture system. LN521 works with most commercial media and hPSCs can routinely be cultured as single cells without the need to add apoptosis inhibitors, such as ROCKi. Since laminin efficiently buffers endogenously produced growth factors, LN521 also performs well together with growth-factor reduced media (Rodin, 2014a). The biorelevant support provided by the LN521 matrix also has 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 workload, time, and cost for growing your cells.
Biolaminin 521 supports clonal culture
With support from the LN521 matrix, cells can be passaged in very low cell densities and can be cultured to high confluence without phenotypic alterations (Rodin, 2014a; Rodin, 2014b). LN521 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 LN521 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 LN521 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 troubleshooting guide (Rodin, 2014b).
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WHAT OUR CUSTOMERS SAY
“Biolaminin 521 has simplified the day-to-day expansion of our human pluripotent stem cells and significantly reduced the operator time. Our new costs are only a fraction of what they used to be when we relied on MEFs.”
Dr. Roger E. Rönn,
Univ. of Edinburgh, Scotland
Biolaminin Key Advantages
Biolaminin 521 LN (LN521) has a positive effect on stabilizing and homogenizing pluripotent gene expression profiles between hES cell lines and provides more controllable, robust, and clinically safe culture conditions. hES cells cultured on Biolaminin 521 are more homogenous, attached better, and grew faster compared to hES cells cultured on other matrices.
Specific laminin isoforms are present in different tissue microenvironments and they are essential for cell survival, proliferation and differentiation. Biolaminin products allow you to imitate the natural cell-matrix interactions in vitro.
All our matrices are chemically defined and animal origin-free, which makes them ideal substrates for each level of the scientific process – from basic research to clinical applications.
Our products have consistent composition and quality. This enables minimized variability between experiments and uniform pluripotency gene expression profiles between different cell lines.
Numerous scientists have found our products and finally succeeded in their specific stem cell application. The power of full-length laminins incorporated into various cell systems is well documented in scientific articles and clinical trials.
Biolaminin 521 LN (LN521)
Human recombinant laminin 521Biolaminin 521 LN is the natural laminin for pluripotent stem cells and therefore reliably facilitates self-renewal of human ES and iPS cells in a chemically defined, feeder-free and animal origin-free stem cell culture system. LN521 is animal origin-free to the primary level.VIEW product
Biolaminin 521 MX (MX521)
Human recombinant laminin 521Biolaminin 521 MX is the natural laminin for pluripotent stem cells and therefore reliably facilitates self-renewal of human ES and iPS cells in a chemically defined, feeder-free stem cell culture system. MX521 is animal origin-free to the secondary level.VIEW product
Biolaminin 521 CTG (CT521)
Human recombinant laminin 521Biolaminin 521 CTG is a full-length, human, recombinant laminin 521 substrate, the only one of its kind on the market, providing an optimal environment for feeder-free culture of human PSCs, MSCs and most anchorage-dependent progenitor cell types. CT521 is animal origin-free to the secondary level and designed for clinical studies.VIEW product
3D culture substrateBiosilk 521 is a natural biomaterial made of spider silk and laminin 521 – an ideal 3D culture substrate for integration, expansion, and long-term differentiation of human pluripotent stem cells and different progenitor cells.VIEW product
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