Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai, China.
Department of Plastic and Reconstructive Surgery, Shanghai General Hospital, Shanghai Jiao Tong University, School of Medicine, Shanghai, China.
Anal Cell Pathol (Amst). 2019 Dec 19;2019:2107414. doi: 10.1155/2019/2107414. eCollection 2019.
Since the first report on the immunomodulatory and immunosuppressive properties of Adipose-Derived Stem Cells (ADSCs), many studies have elucidated the underlying molecular mechanism of their suppressive activity on mixed lymphocyte reaction (MLR). However, a gap exists in our understanding of the molecular mechanism of ADSC-conditioned medium (ADSC-CM) on MLR.
ADSCs were isolated from Human Adipose Tissues, and Enzyme-linked Immunosorbent Assay (ELISA) was used to identify the concentration of transforming growth factor 1 (TGF-1) in ADSC-CM. The transcript abundance of TGF-1, as well as that of insulin-like growth factor binding protein 3 (IGF-BP3), was evaluated using qRT-PCR on Jurkat cells cultured in ADSC-CM for 24 hours. The proliferation of the Jurkat cells was assessed using cell cycle assay. Western blotting was performed to identify potential signaling molecules involved in the ADSC-CM-induced inhibition of Jurkat cell proliferation.
The findings confirm that the isolated ADSCs demonstrate classic ADSC characteristics. The level of TGF-1 was found to be low in ADSC-CM, as assessed by ELISA. Jurkat cells grown in ADSC-CM show reduced gene expression of TGF-1 and IGF-BP3 compared with that of the control group. Furthermore, western blotting of ADSC-CM grown Jurkat cells that were blocked at the G0/G1 stage indicates that ADSC-CM decreases the protein expression of pP38 in a dose-dependent manner.
ADSC-CM can inhibit Jurkat cell proliferation through the TGF-1-p38 signaling pathway.
自脂肪来源干细胞（ADSCs）的免疫调节和免疫抑制特性的首次报告以来，许多研究已经阐明了其对混合淋巴细胞反应（MLR）抑制活性的潜在分子机制。然而，我们对 ADSC 条件培养基（ADSC-CM）对 MLR 的分子机制的理解存在差距。
从人脂肪组织中分离 ADSC，并使用酶联免疫吸附测定（ELISA）来鉴定 ADSC-CM 中转化生长因子 1（TGF-1）的浓度。使用 qRT-PCR 评估 Jurkat 细胞在 ADSC-CM 中培养 24 小时后 TGF-1 和胰岛素样生长因子结合蛋白 3（IGF-BP3）的转录丰度。通过细胞周期测定评估 Jurkat 细胞的增殖。进行 Western blot 以鉴定 ADSC-CM 诱导 Jurkat 细胞增殖抑制中涉及的潜在信号分子。
研究结果证实，分离的 ADSC 表现出典型的 ADSC 特征。通过 ELISA 评估，发现 ADSC-CM 中的 TGF-1 水平较低。与对照组相比，在 ADSC-CM 中生长的 Jurkat 细胞的 TGF-1 和 IGF-BP3 基因表达降低。此外，在 ADSC-CM 中生长的 Jurkat 细胞中阻断 G0/G1 期的 Western blot 表明，ADSC-CM 以剂量依赖性方式降低 pP38 的蛋白表达。
ADSC-CM 可以通过 TGF-1-p38 信号通路抑制 Jurkat 细胞增殖。
