hiPSC Plate coating (Geltrex vs. Laminin)

[zora_b...@hotmail.com]

Hi Abs and the whole TooT community, 

Our lab cultures the hiPSC routinely on Laminin-521. As you use Geltrex (and the coating can have an effect on the lineage commitment), I tried switching the coating to Geltrex for 2-3 passages before starting a differentiation.

However, the different hiPSC lines react super heterogeneously to the change in coating (some love and others dislike it), and it does not really normalize across passages (4+). 

Thus, my question is if you ever compared differentiation capacities (for BMO or the comBO protocol) from hiPSC coming from other coated plate types? 

If not, how essential do you think Geltrex is? 

Thank you so much for generating this group and I look forward to hearing/learning from you. 

Cheers,

Zora

[Abdullah Khan]

Hey Zora ! We haven't tried comparing differentiation capacities, but laminin is well documented as being a good substrate for maintaining iPSCs. We tend not to have too much trouble with the transition to GelTrex (which is a lot cheaper!) If your cells are happy and stable in laminin, then it's worth trying the differentiation. There's a small chance you may have issues, but to be honest if your karyotype and potency assays look good I don't think you will. 

The alternative is to use ESC qualified reduced growth factor matrigel to coat the plates. This is a bit more nutrient dense than GelTrex and cells that seem to struggle to grow on GelTrex do well on ESC-rGF-Matrigel. We have (and others have) differentiated from this matrix with no issues I believe. 

Hope this helps! 

Abs