The future is fully defined: recombinant fragment E8 of laminin-511 is a viable xenofree alternative to Matrigel for hiPSC culture and differentiation into neurovascular cell types

Abstract

Matrigel remains the gold standard substrate for culture of induced pluripotent stem cells (iPSCs). However, its highly variable composition, animal origin and unpredictable effects on biological activity have been discussed for more than 3 decades. In this study, we explore the use of fragment E8 of recombinant laminin 511, commercially available in form of iMatrix-511, as an alternative to Matrigel for iPSC maintenance and differentiation. Female iMR90-4 human iPSCs were cultured on either iMatrix or Matrigel and assessed for cell growth and viability, pluripotency, genetic stability, and ability to differentiate into isogenic brain microvascular endothelial cells (iBMECs) and brain pericytes. It was observed that iMatrix facilitated iPSC growth and viability comparable to Matrigel while maintaining a higher number of more consistently sized colonies. Additionally, like Matrigel, iMatrix maintained the expression of pluripotency markers SSEA-4 and OCT-3/4 over 15 passages without inducing DNA damage. iMatrix also supported the differentiation of these iPSCs into isogenic iBMECs and pericytes, which were successfully co-culture for generation of a simplified blood-brain barrier model. Overall, we showed that iMatrix, which is a cost effective, fully defined, and xenofree alternative can be used as a substitute for Matrigel for maintenance and differentiation of iPSCs.