María Nelcy Orozco García1,
Manuel J. Gayoso1, Sara Gayoso del Villar1, Girish K.
Srivastava1, Manuel Garrosa1
1. Departamento de Biología Celular,
Histología y Farmacología, Facultad de Medicina, Universidad de Valladolid,
Valladolid, Spain.
Abstract
Schwann cell (SC) cultures are required for neural
regeneration, as for example in injured nerves. However, SC culture procedures
need improvements. This study evaluates and compares growth of SC cultures
without and with neuregulin, and in co-cultures with adipose mesenchymal stem
cells (ASCs). Rat originated SCs and ASCs were cultured, maintained and
subcultured to prepare monocultures and co-cultures for this study.
Morphological, immunohistochemical and quantitative evaluations were performed
at 48 and 96 h. In monocultures SCs were S100 and vimentin positive, appeared
bipolar with small somata containing small and elongated nuclei, and tended to
contact on their ends forming chains or clustering around other cell types.
ASCs were CD 105 and vimentin positive, showed fibroblast-like shape with large
rounded nuclei and distinct nucleoli, became larger along time and contacted
with the neighboring cells. In SC-ASC co-culture, a few ASCs appeared in
triangular shape, their CD105 positivity turned weaker and became vimentin
negative at 4 days confirming ASC differentiation initiation towards SC.
Fibroblasts appeared in a small proportion in each culture. Cell counting and
further ANOVA analysis revealed significant differences for cell growth between
48 and 96 h in each culture, whereas across different cultures, significant
differences were found only between SC cultures grown without neuregulin and
rest of SC cultures. In co-cultures ASC proliferated faster. Neuregulin
increases SC cultures growth about 4 times faster than SC cultures alone. A
similar SC proliferation increase was observed in co-cultures of SC with ASC at
96 h, suggesting possible role of ASC released growth factors in SC growth.
Keywords: Nerve Regeneration, Schwann,
Adipose-derived mesenchymal stem cells, Neuregulin, Co-culture
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Publication
Ms. Ref. No.:A0101002
Reviewer 1 Reviewer 2
Received: 01072016
Accepted: 30112016
Article in Press: 15122016
Article in Press: 15122016