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The combined effects of three‐dimensional cell culture and natural tissue extract on neural differentiation of P19 embryonal carcinoma stem cells
Author(s) -
Azizi Faezeh,
Jalil Hamidreza,
Nasiri Zohreh,
Moshtaghian Jamal,
Esmaeili Fariba,
Doostmohammadi Ali,
Shabani Leila,
Ebrahimie Esmaeil
Publication year - 2018
Publication title -
journal of tissue engineering and regenerative medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.835
H-Index - 72
eISSN - 1932-7005
pISSN - 1932-6254
DOI - 10.1002/term.2712
Subject(s) - p19 cell , embryonic stem cell , microbiology and biotechnology , neuroepithelial cell , stem cell , extracellular matrix , neural stem cell , synaptophysin , cellular differentiation , embryonal carcinoma , biology , nestin , cell culture , chemistry , pathology , adult stem cell , immunology , immunohistochemistry , biochemistry , medicine , genetics , gene
Tissue engineering, as a novel transplantation therapy, aims to create biomaterial scaffolds resembling the extracellular matrix in order to regenerate the damaged tissues. Adding bioactive factors to the scaffold would improve cell–tissue interactions. In this study, the effect of chitosan polyvinyl alcohol nanofibres containing carbon nanotube scaffold with or without active bioglass (BG + /BG − ), in combination with neonatal rat brain extract on cell viability, proliferation, and neural differentiation of P19 embryonic carcinoma stem cells was investigated. To induce differentiation, the cells were cultured in α‐MEM supplemented with neonatal rat brain extract on the scaffolds. The expression of undifferentiated stem cell markers as well as neuroepithelial and neural‐specific markers was evaluated and confirmed by real‐time Reverse transcription polymerase chain reaction (RT‐PCR) and immunofluorescence procedures. Finally, the three‐dimensional (3D) cultured cells were implanted into the damaged neural tubes of chick embryos, and their fates were followed in ovo . Based on the histological and immunofluorescence observations, the transplanted cells were able to survive, migrate, and penetrate into the host embryonic tissues. Gene network analysis suggested the possible involvement of neurotransmitters as a downstream target of synaptophysin and tyrosine hydroxylase . Overall, the results of this study indicated that combining the effects of 3D cell culture and natural brain tissue extract can accelerate the differentiation of P19 embryonic carcinoma cells into neuronal phenotype cells.