Standardization of human-induced pluripotent stem cells by optimizing culture conditions and monitoring key parameters
Listed in
This article is not in any list yet, why not save it to one of your lists.Abstract
The differentiation potential of human-induced pluripotent stem cells (hiPSCs) is not constant among clones, therefore standardization of iPSCs as a starting material is a key issue for the dissemination of hiPSC-based cell therapy. We showed that hiPSCs cultured with medium that supported the glycolytic pathway demonstrated a high level of chromodomain-helicase-DNA-binding protein 7 (CHD7) expression and retained differentiation potential. Notably, hiPSCs cultured with medium that supported mitochondrial activity expressed a low level of CHD7 and lost differentiation potential. In addition, we showed secretion of kynurenine by undifferentiated hiPSCs, and 2-AAA when differentiated. Thus, we propose re-cloning iPSC clones with medium supporting the glycolytic pathway for standardization, and measuring the expression of CHD7 by flow cytometry to check the differentiation potential and measuring Kynurenine and 2-AAA levels in the culture medium to monitor and assure the differentiation status of cells during culture.