The use of defined conditions for derivation, maintenance, and differentiation of human-induced pluripotent stem cells (hiPSCs) provides a superior experimental platform to discover culture responses to differentiation cues and elucidate the basic requirements for cell differentiation and fate restriction. Adoption of defined systems for reprogramming, undifferentiated growth, and differentiation of hiPSCs was found to significantly influence early stage differentiation signaling requirements and temporal kinetics for the production of primitive neuroectoderm. The bone morphogenic protein receptor agonist LDN-193189 was found to be necessary and sufficient for neural induction in a monolayer system with landmark antigens paired box 6 and sex-determining region Y-box 1 appearing within 72 h. Preliminary evidence suggests this neuroepithelium was further differentiated to generate ventral spinal neural progenitors that produced electrophysiologically active neurons in vitro, maintaining viability posttransplantation in an immunocompromised host. Our findings support current developments in the field, demonstrating that adoption of defined reagents for the culture and manipulation of pluripotent stem cells is advantages in terms of simplification and acceleration of differentiation protocols, which will be critical for future clinical translation.
Bibliographical noteFunding Information:
The author(s) disclosed receipt of the following financial support for the research and/or authorship of this article: The Wings for Life Spinal Cord Research Foundation, private philanthropy through the University of Minnesota Foundation (UMF), and the KL2 Scholar Program (A. M. Parr) from the Clinical and Translational Science Institute KL2TR113 (NIH:8UL1TR000114).
- human-induced pluripotent stem cells
- neural differentiation
- ventral spinal neurons