Comparison of different protocols for neural differentiation of human induced pluripotent stem cells.
Although embryonic stem cells (ESCs) have enormous potentials due to their pluripotency, their therapeutic use is limited by ethical, biological and safety issues. Compared to ESCs, induced pluripotent stem cells (iPSCs) can be obtained from mouse or human fibroblasts by reprogramming. Numerous studies have established many protocols for differentiation of human iPSCs (hiPSCs) into neural lineages. However, the low differentiation efficiency of such protocols motivates researchers to design new protocols for high yield differentiation. Herein, we compared neural differentiation potential of three induction media for conversion of hiPSCs into neural lineages. In this study, hiPSCs-derived embryoid bodies were plated on laminin coated dishes and were treated with three induction media including (1) bFGF, EGF (2) RA and (3) forskolin, IBMX. Immunofluorescence staining and quantitative real-time PCR (qPCR) analysis were used to detect the expression of neural genes and proteins. qPCR analysis showed that the expression of neural genes in differentiated hiPSCs in forskolin, IBMX supplemented media was significantly higher than undifferentiated cells and those in induction media containing bFGF, EGF or RA. In conclusion, our results indicated a successful establishment protocol with high efficiency for differentiation of hiPSCs into neural lineages.
Department of Nanobiotechnology, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran.
Ali
A
Salimi
Samad
S
Nadri
Marzieh
M
Ghollasi
Khosro
K
Khajeh
Masoud
M
Soleimani
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D004815:Epidermal Growth Factor / Q000008:administration & dosage; D016222:Fibroblast Growth Factor 2 / Q000008:administration & dosage; D005347:Fibroblasts / Q000187:drug effects; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D058953:Neural Stem Cells / Q000166:cytology* / Q000187:drug effects; D014212:Tretinoin / Q000008:administration & dosage
D016222:Fibroblast Growth Factor 2; D014212:Tretinoin; D004815:Epidermal Growth Factor