Laminin-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 (LN-111) is mostly expressed in extra-embryonic basement membranes and stimulates cells to commit to differentiation. Contrary, laminin-521 (LN-521) and laminin-511 (LN-511) are expressed around the stem cells in the inner cell mass and support survival and self-renewal of the pluripotent stem cells (Domogatskaya et al., 2012; Ekblom et al., 2003; Miner & Yurchenco, 2004). 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 et al., 2015). Due to differences in cellular receptor binding activities, LN-521 supports faster hPSC expansion of human ES and iPS cells compared to LN-511 (Rodin et al., 2014).
Human recombinant LN-521 is a chemically defined and xeno-free system for robust, long-term expansion of human pluripotent stem cells (Rodin et al., 2014a; Rodin et al., 2014b; Tano et al., 2014). LN-521 successfully replicates the genuine human stem cell niche in vitro (Rodin et al., 2014a; Laperle et al., 2015) and thereby provides a highly reliable and reproducible culture system to create sufficient populations of undifferentiated cells that, in turn, can be terminally differentiated.
α6β1 integrin mediating effects of LN-521 promotes hPSC survival and self-renewal
LN-521 has strong interaction to the α6β1 integrin which induces the PI3K/Akt signaling pathway. In pluripotent stem cells this interaction promotes survival and stimulating cell proliferation over differentiation (Rodin et al., 2014a). 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 et al., 2016). LN-511 binds the same integrin but the α6β1 integrin mediating effects of LN-521 is much stronger than that of LN-511 which results in more robust hPSC expansion on LN-521 (Rodin et al., 2014).
Robust, long-term expansion of human ES and iPS cells on LN-521
The cells migrate quickly on LN-521, which correlates with the high survival of the cells (Rodin et al., 2014a). LN-521 also support high post-thawed survival of cryopreserved pluripotent stem cells (Miyazaki et al., 2014). Cells grow as a monolayer providing you with a homogenous, pluripotent cell culture without need to dispose of unwanted differentiated cells (Rodin et al., 2014a; Rodin et al., 2014b). Cells amplify faster compared to other matrices, with about 10-fold expansion in 4 days.
With the support from the LN-521 matrix, cells can be passaged in very low cell densities and can be cultured to high confluence without phenotypic alterations (Rodin et al., 2014a). Due to the authentic biorelevant signals are provided by the culture matrice, LN-521 works with most commercial medias. Since laminin efficiently buffers endogenously produced growth factors LN-521 also performs well together with growth-factor reduced medias (Rodin et al., 2014a). The biorelevant support provided by the LN-521 matrix also have a protective effect against DNA damage caused by suboptimal culture conditions, such as high-density culture and pH changes (Jacobs et al., 2016). This allows for weekend-free feeding, drastically reducing work load, time and cost for growing your cells.
LN-521 also supports efficient clonal culture (Rodin et al., 2014a; Tano et al., 2014). Clinically compliant human embryonic stem cell lines can even be derived from a single blastomere (Damdimopoulou et al., 2015; Rodin et al., 2014a; Hovatta et al., 2014) and there is no longer any need to destroy the embryo.
The main benefits of LN-521 for human pluripotent stem cell cultures are:
- Biologically relevant
Natural environment for cultured human ESCs and iPSCs
- Chemically defined and xeno-free
Clinical relevance and less experimental variation
- Rapid cell expansion without spontaneous differentiation
Fast scale-up with about 10-fold expansion in 4 days
- Easy coating and single-cell split/seeding protocol
Standardized monolayer culture system for automation
- Genetically stable cells
High quality, genetically stable cells with maintained phenotype for over 130 passages
- Defined clonal hPSC derivation
Efficient derivation of iPSC and clinical grade human ES cell lines
- Weekend-free feeding
No weekend work and reduced cost
- No lot-to-lot variation
Reliable experimental data
- Medium independent
Works with most commercial media for full flexibility and ease of use
- Scientifically proven in high-impact journals
- Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment. Rodin et al. Nat Commun., 2014a
- Monolayer culturing and cloning of human pluripotent stem cells on laminin-521–based matrices under xeno-free and chemically defined conditions. Rodin et al. Nature Protocols, 2014b
- Functional diversity of laminins. Domogatskaya et al. Annu Rev Cell Dev Biol., 2012
- Expression and biological role of laminin-1. Ekblom et al. Matrix Biol., 2003
- Laminin functions in tissue morphogenesis. Miner & Yurchenco. Annu Rev Cell Dev Biol., 2004
- α-5 Laminin Synthesized by Human Pluripotent Stem Cells Promotes Self-Renewal. Laperle et al. Stem Cell Reports, 2015
- Optimization of slow cooling cryopreservation for human pluripotent stem cells. Miyazaki et al. Genesis, 2014
- Inhibition of FAK Signaling by Integrin α6β1 Supports Human Pluripotent Stem Cell Self‐Renewal. Villa-Diaz et al. Stem Cells, 2016
- Higher-Density Culture in Human Embryonic Stem Cells Results in DNA Damage and Genome Instability. Jacobs et al. Stem Cell Reports, 2016
- A Novel in vitro method for detecting undifferentiated human pluripotent stem cells as impurities in cell therapy products using a highly efficient culture system. Tano et al. PLOS One, 2014
- Animal Substance-Free Human Embryonic Stem Cells Aiming at Clinical Applications. Hovatta et al. Stem Cells Transl Med. 2014
- Human embryonic stem cells. Damdimopoulou et al.