Anat Cell Biol.  2015 Sep;48(3):170-176. 10.5115/acb.2015.48.3.170.

Differentiation of adipose-derived stem cells into Schwann-like cells: fetal bovine serum or human serum?

• Younesi E
• Bayati V
• Hashemitabar M
• Azandeh SS
• Bijannejad D
• Bahreini A

• KMID: 2424819
• DOI: http://doi.org/10.5115/acb.2015.48.3.170

Abstract

Access to autologous Schwann cells is limited due to lack of donor site and its difficult isolation and culture. Therefore, one of the possible ways to obtain to Schwann cells is to differentiate mesenchymal stem cells into glial pathway using various materials and protocols. The aim of this study was to compare the effects of fetal bovine serum and human serum on Schwann cell differentiation of adipose-derived stem cells to choose the best serum for use in future research. For this purpose, after isolation of human adipose-derived stem cells, it was characterized and differentiated into Schwann cell lineage using two protocols which one of them contained fetal bovine serum and the other human serum. At the end, morphological evaluation declared an increased detachment of cells in response to human serum. On the other side, immunocytochemistry showed that there was a significant increase in the number of cells expressing glial fibrillary acidic proteins and S100 in fetal bovine serum-treated group when compared to human serum-treated one (P<0.05). It was concluded that fetal bovine serum was more effective than allogeneic human serum in Schwann cell differentiation of adipose-derived stem cells.

Keyword

Adipose-derived stem cells; Schwann cell; Differentiation; Human serum

MeSH Terms

Cell Differentiation
Cell Lineage
Glial Fibrillary Acidic Protein
Humans*
Immunohistochemistry
Mesenchymal Stromal Cells
Schwann Cells
Stem Cells*
Tissue Donors
Glial Fibrillary Acidic Protein

Figure

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  Fig. 1 Adipose-derived stem cells (ASCs) isolation and culture. (A) Cultured cells of stromal vascular fraction of fat tissue on day 3 of primary culture. (B) ASCs at passage 2 at confluence. Scale bars=30 µm (A, B).

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  Fig. 2 Differentiation potential assay of adipose-derived stem cells (ASCs) at fourth passage. (A) Lipid vacuoles inside the cytoplasm of adipoblasts appear as red circular spots after staining with oil Red O which stains triglyceride and neutral lipid. (B) Calcium deposition was shown by alizarin red S staining as dark red spots in the plate. Scale bars=30 µm (A, B).

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  Fig. 3 Morphological assessment of Schwann-like (SC-like) cells after 14 days of induction. (A) Adipose-derived stem cells at fourth passage on day 0 of induction by β-mercaptoethanol. (B) SC-like cells with deformed morphologies in human serum (HS)-treated groups. Most of cells detached in response to the differentiation medium containing allogeneic HS. (C) SC-like cells appeared mostly bipolar or tripolar, typical morphology of Schwann cells when exposed to the differentiation medium containing fetal bovine serum. Scale bars=40 µm (A-C).

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  Fig. 4 Immunofluorescence staining indicated that the differentiated Schwann-like cells expressed S100 (A, C) and glial fibrillary acidic protein (GFAP) (B, D) in both human serum (HS)- and fetal bovine serum (FBS)-treated groups. Fibroblasts 3T3 and undifferentiated adipose-derived stem cells were taken as negative control and new born rat Schwann cells was considered as positive control. Scale bars=30 mm (A-D).

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  Fig. 5 Quantitative analysis of cells expressing glial fibrillary acidic protein (GFAP) (A) and S100 (B) in both fetal bovine serum (FBS)- and human serum (HS)-treated groups (data are mean cells±SEM, %). *P<0.05.

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