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Derivation of motor neuron‐like cells from neonatal mouse testis in a simple culture condition
Author(s) -
Javanmardy S.,
Asadi M. H.,
Movahedin M.,
Moradpour F.,
Bahadoran H.
Publication year - 2016
Publication title -
andrologia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.633
H-Index - 59
eISSN - 1439-0272
pISSN - 0303-4569
DOI - 10.1111/and.12545
Subject(s) - nestin , embryoid body , biology , neural stem cell , stem cell , embryonic stem cell , sox2 , microbiology and biotechnology , motor neuron , amniotic epithelial cells , retinoic acid , neurogenesis , neuron , sonic hedgehog , immunology , adult stem cell , neuroscience , cell culture , biochemistry , genetics , gene , spinal cord , signal transduction
Summary Embryonic stem cell ( ESC ) therapy is an exciting way to treat neurodegenerative disease and central nervous system injury. However, many ethical and immunological problems surround the use of embryonic stem cells. Finding an alternative source of stem cells is therefore pertinent. In this study, spermatogonia stem cells ( SSC s) were used to generate mature motor neurons. SSC s were extracted from neonatal testes and cultured in DMED /F12 medium for 3 weeks. Characterisation of SSC ‐derived ESC ‐like cells was confirmed by RT ‐ qPCR , immunostaining, alkaline phosphatase activity and their ability to form embryoid bodies ( EB s). The EB s were induced by retinoic acid and Sonic hedgehog and trypsinised to obtain single induced cells. The single cells were cultured in neural medium for 18 days. Characterisation of neural precursors and motor neuron‐like cells was confirmed by RT ‐ qPCR and immunocytochemical analysis at the 7th day (early stage) and 18th day (late stage), respectively, of culturing. The neural precursors were found to be positive for nestin and Sox2, and a small fraction of cells expressed β‐tubulin III . Upon further differentiation, multipolar neurons were detected that expressed β‐tubulin III and MAP 2 markers. Moreover, the expression levels of Olig2 and PAX 6 were significantly lower, while HB 9, Isl1 and Isl2 expression levels were higher at the late stage when compared to the early stage. These results show that SSC s have the potential to differentiate to motor neuron‐like cells and express markers specific for mature motor neurons. However, the functional ability of these cells remains to be evaluated in future studies.