Open AccessSpecific Marker Expression and Cell State of Schwann Cells during Culture In Vitro
Author(s) -
Zhangyin Liu,
Yuqing Jin,
Lulu Chen,
Yang Wang,
Xiaonan Yang,
Jing Cheng,
Wei Wu,
Zuoliang Qi,
Zhe Shen
Publication year - 2015
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0123278
Subject(s) - sox10 , sox2 , gap 43 protein , biology , schwann cell , western blot , microbiology and biotechnology , myelin basic protein , immunofluorescence , in vitro , in vivo , stem cell , immunohistochemistry , immunology , transcription factor , embryo , central nervous system , antibody , gene , myelin , genetics , neuroscience , neural crest
Schwann cells (SCs) in animals exist in different developmental stages or wound repair phases, distinguished mainly by the expression of SC-specific markers. No study has yet determined SC state under in vitro culture conditions, and the specific markers expressed in SC are obscure as well. In this study, we harvested sciatic nerves from newborn mice and isolated SCs by an enzyme-digestion method, then we examined the expression profiles of ten markers (S100, p75 NTR , Sox10, Sox2, GAP43, NCAM, Krox20, Oct6, MBP, and MPZ) at both the RNA and protein levels in in vitro mouse SCs and speculated their relation with in vivo SC stages. We assayed RNA and protein levels of SC specific markers by immunofluorescence, Western Blot, and real-time quantitative RT-PCR. The results show that the expression of most markers (S100, p75 NTR , GAP43, NCAM, Krox20, Oct6, MBP and MPZ) was not detectable in all of early stage cultured SCs. The expression of transcription factors Sox10 and Sox2 was, however, detectable in all SCs. After 8 days, the positive expression rate of all markers except GAP43 and Oct6 was almost 100%.These results indicates Sox10 is a necessary marker for SC identification, while S100 is not reliable. SCs cultured in vitro express Sox2, P75 NTR , NCAM, GAP43, Oct6, and MPZ, suggesting that they are similar to in vivo undifferentiated iSCs or dedifferentiated iSCs after nerve injury.