Combination of basic fibroblast growth factor and epidermal growth factor enhances proliferation and neuronal/glial differential of postnatal human enteric neurosphere cells in vitro.
Human enteric neural stem cells (hENSCs) proliferate and differentiate into neurons and glial cells in response to a complex network of neurotrophic factors to form the enteric nervous system. The primary aim of this study was to determine the effect of basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) on in-vitro expansion and differentiation of postnatal hENSCs-containing enteric neurosphere cells. Enteric neurosphere cells were isolated from rectal polyp specimens of 75 children (age, 1-13 years) and conditioned with bFGF, EGF, bFGF+EGF, or plain culture media. Proliferation of enteric neurosphere cells was examined using the methyl thiazolyl tetrazolium colorimetric assay over 7 days of culture. Fetal bovine serum (10%) was added to induce the differentiation of parental enteric neurosphere cells, and differentiated offspring cells were immunophenotyped against p75 neutrophin receptor (neural stem cells), peripherin (neuronal cells), and glial fibrillary acidic protein (glial cells). Combining bFGF and EGF significantly improved the proliferation of enteric neurosphere cells compared with bFGF or EGF alone (both P<0.01) throughout 7 days of culture. The addition of bFGF drove a significantly greater proportion of enteric neurosphere cells to differentiate into neuronal cells than that of EGF (P<0.01), whereas addition of EGF resulted in significantly more glial differentiation compared with addition of bFGF (P<0.01). Combining bFGF and EGF drove enteric neurosphere cells to differentiate into neuronal cells in a proportion similar to glial cells. Our results showed that the combination of bFGF and EGF significantly enhanced the proliferation and differentiation of postnatal hENSCs-containing enteric neurosphere cells in vitro.
aDepartment of Pediatric Surgery, The Second Affiliated Hospital, Xi'an Jiaotong University bMinistry of Education Key Laboratory of Environment and Disease Related Genetics, Xi'an Jiaotong University College of Medicine, Xi'an cEndoscopy Center, Shaanxi 215 Nuclear Industry Hospital, Xianyang, China.
Wei-Kang
WK
Pan
Hui
H
Yu
A-Li
AL
Wu
Ya
Y
Gao
Bai-Jun
BJ
Zheng
Peng
P
Li
Wei-Li
WL
Yang
Qiang
Q
Huang
Huai-Jie
HJ
Wang
Xin
X
Ge
D000293:Adolescent; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002648:Child; D002675:Child, Preschool; D003124:Colorimetry; D017615:Enteric Nervous System / Q000166:cytology*; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005260:Female; D005904:Glial Fibrillary Acidic Protein / Q000494:pharmacology*; D006801:Humans; D007223:Infant; D008297:Male; D009419:Nerve Tissue Proteins / Q000378:metabolism; D058953:Neural Stem Cells / Q000187:drug effects*; D009457:Neuroglia / Q000187:drug effects* / Q000378:metabolism; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D017475:Receptors, Nerve Growth Factor / Q000378:metabolism
D005904:Glial Fibrillary Acidic Protein; C484065:NGFR protein, human; D009419:Nerve Tissue Proteins; D017475:Receptors, Nerve Growth Factor; D004815:Epidermal Growth Factor