The laminin protein family is conserved
The laminin proteins of the extracellular membrane are highly conserved and human recombinant laminins can be used for defined and reliable culture of many animal origin pluripotent stem cells (Domogatskaya et al., 2012).
Mouse Pluripotent Stem Cells
In contrast to the inner cell mass of the human embryo where both laminin-521 (LN-521) and laminin-511 (LN-511) is expressed, the only laminin isoform expressed in the early mouse embryo is LN-511. This data is nicely presented in an article published in Nature Cell Biology by the groups of Drs. Austin Smith and Jennifer Nichols where they also show that laminin-511 is crucial for acquisition of naive pluripotency (Boroviak et al., 2014).
Since human recombinant LN-511 provides the mouse PSC with their natural environment it supports long-term expansion of mouse embryonic stem cells (Domogatskaya et al., 2008). LN-511 alone, without the addition of leukemia inhibitory factor (LIF) or other differentiation inhibitors, is sufficient to enable self-renewal of mouse ES cells for up to 169 days (31 passages) with maintained expression of pluripotency markers. Cells cultured for 95 days (17 passages) on LN-511 were used to successfully generate chimeric mice (Domogatskaya et al., 2008). This data is supported by a publication by Lecht and co-coworkers where they show that laminin is one of the major pro-adhesive ECM protein for mESCs. The mESCs were found to adhere with highest potencies to LN-511 and LN-521, most likely via specific binding to a3B1 and a6B1 integrin receptors expressed on the mES cell surface (Lecht et al., 2013).
The main benefits of LN-511 for mouse pluripotent stem cells are:
- Fast adherence of mESCs to LN-511
3 to 5-fold higher affinity than other matrices
- Rapid cell expansion with minimal spontaneous differentiation
Reliable and fast scale-up where cells grow as a homogenous monolayer
- Biorelevant matrix where no differentiation inhibitors are needed
Self-renewal of mouse ES cells for over 5 months without the presence of LIF
- Highly efficient single-cell mESC derivation
LN-511 is crucial for acquisition of naive pluripotency
- Chemically defined
No lot-to-lot variation for less experimental variation
- Easy coating and single-cell split/seeding protocol
Standardized monolayer culture system for automation
- Scientifically proven in high-impact journals
Monkey Pluripotent Stem Cells
Laminin-521 and laminin-511 are two of the first extra cellular proteins expressed during development. Both laminin isoforms are present in the inner cell mass of the human embryo and expressed by the pluripotent cells (Domogatskaya et al., 2012).
Human recombinant laminin-521 is a chemically defined and xeno-free system for robust, long-term expansion of human pluripotent stem cells. Since it successfully replicates the specific human stem cell niche in vitro, it makes it the optimal matrix for efficient generation and robust large-scale expansion of human ES and iPS cells (Rodin et al. 2014). Several researchers are successfully routinely using LN-521 for pluripotent expansion of monkey PSC (Unpublished data).
- The ability of inner-cell-mass cells to self-renew as embryonic stem cells is acquired following epiblast specification. Boroviak et al. Nat Cell Biol., 2014
- Laminin-511 but not -332, -111, or -411 enables mouse embryonic stem cell self-renewal in vitro. Domogatskaya et al. Stem Cells, 2008
- Enhanced reseeding of decellularized rodent lungs with mouse embryonic stem cells. Lecht et al. Biomaterials, 2013
- Functional Diversity of Laminins Domogatskaya et al. Annu Rev Cell Dev Biol., 2012
- Clonal culturing of human embryonic stem cells on laminin-521/E-cadherin matrix in defined and xeno-free environment. Rodin et al., Nat Commun., 2014