Department of Plastic and Reconstructive Surgery, Kansai Medical University, 2-5-1 Shin-machi, Hirakata, Osaka 573-1010, Japan.
Cells. 2019 Oct 8;8(10):1218. doi: 10.3390/cells8101218.
Recently, human adipose-derived stem cells (hASCs) were discovered in the human subcutaneous adipose tissue. PLTMax Human Platelet Lysate (PLTMax), a supplement refined from human platelets, has been reported to have proliferative effects on bone marrow mesenchymal stem cells. The proliferative effects of PLTMax on ASCs were investigated in this study.
The ASCs in DMEM (serum-free), DMEM+PLTMax (1%, 2%, 5%, and 10%), and DMEM+FBS (10%) were cultivated for two, five, and seven days. The cell growth rate was examined, BrdU incorporation, and the cell cycle and Ki-67 immunostaining were performed. The cell growth rate was investigated when each inhibitor (PD98059, SP600125, SB203580, and LY294002) was added and phosphorylation of ERK1/2, JNK, p38, and Akt were examined by western blotting. The cell surface marker of hASCs was also analyzed.
The cells in the PLTMax (5%) group showed significantly more proliferation compared to the cells in control (serum-free) and FBS (10%) groups, and a significant increase in the number of cells in the S phase and G2/M phase. The number of Ki-67 positive cells increased significantly in the DMEM+ PLTMax (5%) and the FBS (10%) groups. The addition of inhibitors PD98059, SP600125, SB203580, and LY294002 decreased the proliferative effects of PLTMax on ASCs. Phosphorylation of ERK1/2, JNK, p38, and Akt was observed in both the PLTMax (5%) and the FBS (10%) groups.
For human adipose stem cells, 5% PLTMax was the optimum concentration, which showed a significantly higher proliferative effect than 10% FBS. PLTMax is a useful medium additive, which can substitute FBS. The proliferative effects of PLTMax are suggested to function via multiple signaling pathways, similar to FBS.
最近,人们在人体皮下脂肪组织中发现了人类脂肪来源的干细胞(hASCs)。PLTMax 人血小板裂解液(PLTMax)是一种从人血小板中提取的、具有增殖作用的补充剂,据报道它对骨髓间充质干细胞有增殖作用。本研究旨在探讨 PLTMax 对 ASCs 的增殖作用。
在 DMEM(无血清)、DMEM+PLTMax(1%、2%、5%和 10%)和 DMEM+FBS(10%)中培养 ASCs 2、5 和 7 天。检测细胞生长率、BrdU 掺入、细胞周期和 Ki-67 免疫染色。加入每种抑制剂(PD98059、SP600125、SB203580 和 LY294002)后,研究细胞生长率,并通过 Western blot 检测 ERK1/2、JNK、p38 和 Akt 的磷酸化。还分析了 hASCs 的细胞表面标志物。
与对照组(无血清)和 FBS(10%)相比,PLTMax(5%)组的细胞增殖显著增加,S 期和 G2/M 期的细胞数量显著增加。DMEM+PLTMax(5%)和 FBS(10%)组中 Ki-67 阳性细胞数量显著增加。加入抑制剂 PD98059、SP600125、SB203580 和 LY294002 后,PLTMax 对 ASCs 的增殖作用降低。PLTMax(5%)和 FBS(10%)组均观察到 ERK1/2、JNK、p38 和 Akt 的磷酸化。
对于人脂肪干细胞,5%PLTMax 是最佳浓度,其增殖效果明显高于 10%FBS。PLTMax 是一种有用的培养基添加剂,可替代 FBS。PLTMax 的增殖作用可能通过多条信号通路发挥作用,与 FBS 相似。