Department of Obstetrics, Gynecology and Reproductive Sciences, Center for Reproductive Sciences, and Center for Reproductive Health, University of California San Francisco, 550 16th Street, 7th Floor, Box 0132, San Francisco, CA, 94158, USA.
J Assist Reprod Genet. 2018 May;35(5):757-770. doi: 10.1007/s10815-018-1130-8. Epub 2018 Feb 5.
The study aims to test the hypothesis that platelet-rich plasma (PRP) stimulates cellular processes involved in endometrial regeneration relevant to clinical management of poor endometrial growth or intrauterine scarring.
Human endometrial stromal fibroblasts (eSF), endometrial mesenchymal stem cells (eMSC), bone marrow-derived mesenchymal stem cells (BM-MSC), and Ishikawa endometrial adenocarcinoma cells (IC) were cultured with/without 5% activated (a) PRP, non-activated (na) PRP, aPPP (platelet-poor-plasma), and naPPP. Treatment effects were evaluated with cell proliferation (WST-1), wound healing, and chemotaxis Transwell migration assays. Mesenchymal-to-epithelial transition (MET) was evaluated by cytokeratin and vimentin expression. Differential gene expression of various markers was analyzed by multiplex Q-PCR.
Activated PRP enhanced migration of all cell types, compared to naPRP, aPPP, naPPP, and vehicle controls, in a time-dependent manner (p < 0.05). The WST-1 assay showed increased stromal and mesenchymal cell proliferation by aPRP vs. naPRP, aPPP, and naPPP (p < 0.05), while IC proliferation was enhanced by aPRP and aPPP (p < 0.05). There was no evidence of MET. Expressions of MMP1, MMP3, MMP7, and MMP26 were increased by aPRP (p < 0.05) in eMSC and eSF. Transcripts for inflammation markers/chemokines were upregulated by aPRP vs. aPPP (p < 0.05) in eMSC and eSF. No difference in estrogen or progesterone receptor mRNAs was observed.
This is the first study evaluating the effect of PRP on different human endometrial cells involved in tissue regeneration. These data provide an initial ex vivo proof of principle for autologous PRP to promote endometrial regeneration in clinical situations with compromised endometrial growth and scarring.
本研究旨在验证血小板富血浆（PRP）刺激与临床管理不良子宫内膜生长或宫腔内瘢痕相关的子宫内膜再生细胞过程这一假说。
将人子宫内膜基质成纤维细胞（eSF）、子宫内膜间充质干细胞（eMSC）、骨髓间充质干细胞（BM-MSC）和 Ishikawa 子宫内膜腺癌细胞（IC）与/或 5%激活（a）PRP、非激活（na）PRP、血小板贫浆（aPPP）和非激活贫浆（naPPP）孵育。通过 WST-1、伤口愈合和趋化性 Transwell 迁移实验评估治疗效果。通过角蛋白和波形蛋白的表达评估间充质向上皮转化（MET）。通过多重 Q-PCR 分析各种标志物的差异基因表达。
与 naPRP、aPPP、naPPP 和载体对照相比，激活的 PRP 以时间依赖的方式增强了所有细胞类型的迁移（p<0.05）。WST-1 试验表明，与 naPRP、aPPP 和 naPPP 相比，aPRP 增强了基质和间充质细胞的增殖（p<0.05），而 IC 增殖则受到 aPRP 和 aPPP 的增强（p<0.05）。没有证据表明存在 MET。aPRP 增加了 eMSC 和 eSF 中 MMP1、MMP3、MMP7 和 MMP26 的表达（p<0.05）。与 aPPP 相比，eMSC 和 eSF 中炎症标志物/趋化因子的转录物被 aPRP 上调（p<0.05）。未观察到雌激素或孕激素受体 mRNA 的差异。
这是第一项评估 PRP 对参与组织再生的不同人子宫内膜细胞的影响的研究。这些数据提供了自体 PRP 促进临床中因子宫内膜生长不良和瘢痕形成而受损的子宫内膜再生的初步离体原理证明。
