Differentiation of human umbilical cord mesenchymal stem cells into steroidogenic cells in vitro

Xing,Xiaoyu; Zhang,Zhiyuan; Zhong,Liang; Ju,Guanqun; Zou,Xiangyu; Zhu,Yingjian; Sun,Jie

doi:10.3892/etm.2016.3815

Differentiation of human umbilical cord mesenchymal stem cells into steroidogenic cells in vitro

Authors:
- Xiaoyu Xing
- Zhiyuan Zhang
- Liang Zhong
- Guanqun Ju
- Xiangyu Zou
- Yingjian Zhu
- Jie Sun
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Affiliations: Department of Urology, Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine, Shanghai 200127, P.R. China, Department of Urology, Shanghai First People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200080, P.R. China
Published online on: October 18, 2016 https://doi.org/10.3892/etm.2016.3815
Pages: 3527-3534
Copyright: © Xing et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Although previous studies have shown that stem cells can be differentiated into Leydig cells by gene transfection, a simple, safe and effective induction method has not yet been reported. Therefore, the present study investigated novel methods for the induction of human umbilical cord mesenchymal stem cell (HUMSC) differentiation into Leydig‑like, steroidogenic cells. HUMSCs were acquired using the tissue block culture attachment method, and the expression of MSC surface markers was evaluated by flow cytometry. Leydig cells were obtained by enzymatic digestion and identified by lineage‑specific markers via immunofluorescence. Third‑passage HUMSCs were cultured with differentiation‑inducing medium (DIM) or Leydig cell‑conditioned medium (LC‑CM), and HUMSCs before induction were used as the control group. Following the induction of HUMSCs, Leydig cell lineage‑specific markers (CYP11A1, CYP17A1 and 3β‑HSD) were positively identified using immunofluorescence analysis. Additionally, reverse transcription‑quantitative polymerase chain reaction and western blot analysis were performed to evaluate the expression levels of these genes and enzymes. In contrast, the control group cells did not show the characteristics of Leydig cells. Collectively, these results indicate that, under in vitro conditions, LC‑CM can achieve a comparable effect to that of DIM on inducing HUMSCs differentiation into steroidogenic cells.

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