Differentiation of Adipose-derived Stem Cells into Schwann Cell Phenotype in Comparison with Bone Marrow Stem Cells

Document Type: Original Article

Authors

1 School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

2 School of Medicine, Yasouj University of Medical Sciences, Yasouj, Iran

Abstract

Objective(s)
Bone marrow is the traditional source of human multipotent mesenchymal stem cells (MSCs), but adipose tissue appears to be an alternative and more readily available source. In this study, rat adipose-derived stem cells (ADSCs) were induced to differentiate into Schwann-like cells and compared with rat bone marrow stem cells (BMSCs) for their Schwann-like cells differentiation potential.
Materials and Methods
BMSCs and ADSCs were characterized for expression of MSCs-specific markers, osteogenic and adipogenic differentiation. They were induced to differentiate into Schwann-like cells and analyzed for expression of the Schwann specific markers. The immunocytochemical differentiation markers were S-100 and real time quantitative Real-time polymerase chain reaction (RT-PCR) markers were S100, P75 and glial fibrillary acidic protein (GFAP). 3-(4, 5-Dimethylthiazol- 2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and Annexin V-Fluorescein isothiocyanate (FITC)/ Propidium iodide (PI) double labeling method were employed to detect early stage cell apoptosis.
Results
BMSCs and ADSCs showed similarities in expression of the MSC-specific markers, osteogenic and adipogenic differentiation. Both quantitative RT-PCR and immunocytochemical analysis demonstrated that BMSCs and ADSCs had equal expression of the Schwann-specific markers following Schwann-like cells differentiation. However, gene expression of P75 was higher in BMSCs compared with ADSCs. MTT assay and flow cytometry found that of the total BMSCs and ADSCs in the culture medium, 20% to 30% of the cells died, but the remaining cell population remained strongly attached to the substrate and differentiated.
Conclusion
Comparative analysis showed that Schwann-like cell differentiation potential of ADSCs was slightly decreased in comparison with BMSCs. Therefore, BMSCs are more favorable choice than ADSCs for tissue engineering.

Keywords


1.Evans GR. Peripheral nerve injury: a review and approach to tissue engineered constructs. Anat Rec 2001; 263:396-404.

2.Kim DH, Connolly SE, Kline DG, Voorhies RM, Smith A, Powell M, Yoes T, Daniloff JK. Labeled Schwann cell transplants versus sural nerve grafts in nerve repair. J Neurosurg 1994; 80:254-260.

3.Mahay DG Terenghi. Schwann cell mediated trophic effects by differentiated mesenchymal stem cells. Exp Cell Res 2008; 15:2692-2701.

4.Lundborg G. Alternatives to autologous nerve grafts. Handchir Mikrochir Plast Chir 2004; 36:1-7.

5.Lin W, Chen X, Wang X, Liu J, Gu X. Adult rat bone marrow stromal cells differentiate into Schwann-like cells in vitro. In Vitro Cell Dev Biol Anim 2008; 44:31-40.

6.Keilhoff G, Stang F, Goihl A, Wolf G, Fansa H. Transdifferentiated mesenchymal stem cells as alternative therapy in supporting nerve regeneration and myelination. Cell Mol Neurobiol 2006; 26:1235-1252.

7.Caddick J, Kingham PJ, Gardiner NJ, Wiberg M, Terenghi G. Phenotypic and functional characteristics of mesenchymal stem cells differentiated along a Schwann cell lineage. Glia 2006; 54:840-849.

8.Dezawa M. Central and peripheral nerve regeneration by transplantation of Schwann cells and transdifferentiated bone marrow stromal cells. Anat Sci Int 2002; 77:12-25.

9.Tohill M, Mantovani C, Wiberg M, Terenghi G. Rat bone marrow mesenchymal stem cells express glial markers and stimulate nerve regeneration. Neurosci Lett 2004; 362:200-203.

10.Kingham PJ, Kalbermatten DF, Mahay D, Armstrong SJ, Wiberg M, Terenghi G. Adipose-derived stem cells differentiate into a Schwann cell phenotype and promote neurite outgrowth in vitro. Exp Neurol 2007; 207:267-274.

11.Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 2002; 13:4279-4295.

12.Strem BM, Hicok KC, Zhu M, Wulur I, Alfonso Z, Schreiber RE, et al. Multipotential differentiation of adiposetissue-derived stem cells. Keio J Med 2005; 54: 132-141.

13.De Ugarte DA, Alfonso Z, Zuk PA, Elbarbary A, Zhu M, Ashjian P, et al. Differential expression of stem cell mobilization-associated molecules on multi-lineage cells from adipose tissue and bone marrow. Immunol Lett 2003; 89: 267-270.

14.Jiang L, Zhu JK, Liu XL, Xiang P, Hu J, Yu WH. Differentiation of rat adipose tissue-derived stem cells into Schwann-like cells in vitro. Neuroreport 2008; 2:1015-1019.

15.Xu Y, Liu Z, Liu L, Zhao C, Xiong F, Zhou C, et al. Neurospheres from rat adipose-derived stem cells could be induced into functional Schwann cell-like cells in vitro. BMC Neurosci 2008; 12:9-21.

16. Tang YJ, Zhang LH, Liu JM, Dong WR, Guo JS, Wang HH, et al. Induced differentiation of rat adipose- derived stem cells into Schwann-like cells. Nan Fang Yi Ke Da Xue Xue Bao 2009; 29:680-684.

17.Zaminy A, Ragerdi Kashani I ,Bafbarestani M, Hedayatpour A, Mahmoudi R, Farzaneh Nejad A. Osteogenic differentiation of rat mesenchymal stem cells from adipose tissue in comparison with bone marrow mesenchymal stem cells: melatonin as a differentiation factor. Iran Biomed J 2008; 12:133-141.

18.Brockes JP, Fields KL, Raff MC.Studies on cultured rat Schwann cells. Establishment of purified populations from cultures of peripheral nerve. Brain Res 1979; 165:105-118.

19.Feller N, Kelder A, Westra G, Ossenkoppele GJ, Schuurhuis GJ. Positive selection for CD90 as a purging option in acute myeloid leukemia stem cell transplants. Cytometry B Clin Cytom 2008; 74:9-16.

20.Lin CS, Xin ZC, Deng CH, Ning H, Lin G, Lue TF.Recent advances in andrology-related stem cell research. Asian J Androl 2008; 10:171-175.

21.Pittenger MF, Mackay AM , Beck SC, Jaiswal RK, Douglas R, Mosca JD, et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284: 143-147. 

22.Leobon B, Roncalli J, Joffre C, Mazo M, Boisson M, Barreau C, et al. Adipose-derived cardiomyogenic cells: in vitro expansion and functional improvement in a mouse model of myocardial infarction. Cardiovasc Res 2009; 1:757-767.

23.Fortino V, Torricelli C, Gardi C, Valacchi G, Rossi Paccani S, Maioli E. ERKs are the point of divergence of PKA and PKC activation by PTHrP in human skin fibroblasts. Cell Mol Life Sci 2002; 59:2165-2171.

24.Jessen KR, Mirsky R Signals that determine Schwann cell identity. J Anat 2002; 200:367-376.