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. 2012 Jul;13(3):151-7.

Derivation of Adipocytes from Human Endometrial Stem Cells (EnSCs)

Jafar Ai  1 , Ahmad Reza Shahverdi, Somayeh Ebrahimi Barough, Homa Mohseni Kouchesfehani, Saeed Heidari, Reza Roozafzoon, Javad Verdi, Ahad Khoshzaban
Affiliations

Derivation of Adipocytes from Human Endometrial Stem Cells (EnSCs)

Jafar Ai et al. J Reprod Infertil. 2012 Jul.

Abstract

Background: Due to increasing clinical demand for adipose tissue, a suitable cell for reconstructive adipose tissue constructs is needed. In this study, we investigated the ability of Human Endometrial-derived stem cells (EnSCs) as a new source of mesenchymal stem cells to differentiate into adipocytes. EnSCs are the abundant and easy available source with no immunological response, for cell replacement therapy.

Methods: Single-cell suspensions of EnSCs were obtained from endometrial tissues from 10 women experiencing normal menstrual cycles, and were cultured at clonal density (10 cells/cm (2) ) or limiting dilution. Endometrial mesenchymal stem cell markers were examined flow cytometry. These cells were treated with adipogenic-inducing medium for 28 days. The adipogenic differentiation of the EnSC was assessed by cellular morphology and further confirmed by Oil Red O staining and RT-PCR. The BM-MSC differentiated into adipocytes in the presence of adipogenic stimuli for 3 weeks.

Results: The flow cytometric analysis showed that the cells were positive for CD90, CD105, CD146 and were negative for CD31, CD34.We showed that the key adipocytes marker PPARa was expressed in mRNA level after 28 days post treatment (PT).

Conclusion: According to our finding, it can be concluded that EnSCs represent a useful in vitro model for human adipogenesis, and provide opportunities to study the stages prior to commitment to the adipocyte lineage.

Keywords: Adipocyte cell; Differentiation; Endometrial stem cell.

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Figures

Figure 1
Figure 1
Morphology of Cultured EnSCs; A: Morphology of freshly isolated EnS cells; B: Fibroblast-like morphology of EnSCs after 2 weeks cell culture; C: Clonal population of EnSCs after plating in 24 well plate 1 week after cloning; D: The same population 2 weeks after cloning. Growth curve for EnSCs after 7 days cell culture (×100 magnification)
Figure 2
Figure 2
Flow cytometric analysis of isolated EnSCs for mesenchymal stem cell markers (CD90, CD105 and CD146), hematopoietic marker (CD34), endothelial marker (CD31). As shown in figure 2 the isolated cells are positive for CD90, CD105 and CD146 and are negative for CD31, CD34
Figure 3
Figure 3
EnSCs differentiate into adipocytes. EnSCs before (A) and after differentiation into adipocytes at 12 days PT (B) and at 21 days PT (C), as demonstrated by light microscopy (A,B,C), Oil Red O staining to demonstrate lipid accumulation at 28 days PT (D,E). Arrows in B and C show adipocyte cells (×100 magnification)
Figure 4
Figure 4
Adipocyte-related gene expression analysis of EnSCs 28 days PT using RT-PCR. β-actin was used as internal standard
See this image and copyright information in PMC

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