脐带间充质干细胞条件培养基作为人颗粒细胞培养基的效果：一项实验研究。

Effect of conditioned medium of umbilical cord-derived mesenchymal stem cells as a culture medium for human granulosa cells: An experimental study.

作者信息

Sumapraja Kanadi, Hestiantoro Andon, Liem Isabella Kurnia, Boediono Arief, Z Jacoeb Teuku

机构信息

Division of Reproductive Immunoendocrinology, Department of Obstetrics and Gynecology, Faculty of Medicine Universitas Indonesia, Dr. Cipto Mangunkusumo Hospital, Jakarta, Indonesia.

Cluster of Human Reproduction, Fertility and Family Planning, the Indonesian Medical Education and Research Institute Universitas Indonesia, Jakarta, Indonesia.

出版信息

Int J Reprod Biomed. 2022 Jan 12;19(12):1037-1044. doi: 10.18502/ijrm.v19i12.10054. eCollection 2021 Dec.

DOI:10.18502/ijrm.v19i12.10054

PMID:35098005

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8792381/

Abstract

BACKGROUND

The umbilical cord-derived mesenchymal stem cells conditioned medium (UC-MSCs-CM) produces secretomes with anti-apoptotic properties, and has the potential to prevent apoptosis of granulosa cells (GC) during controlled ovarian hyperstimulation.

OBJECTIVE

To observe the effect of UC-MSCs-CM on the interaction between pro- and anti-apoptotic proteins and the influence of growth differentiation factor 9 (GDF9) production in GC.

MATERIALS AND METHODS

UC-MSCs-CM was collected from umbilical cord stem cell culture on passage 4. GC from 23 women who underwent in vitro fertilization were cultured and exposed to UC-MSCs-CM for 24 hr. Then RNA of the GC was extracted and the mRNA expression of BCL-2 associated X (BAX), survivin and GDF9 were analysed using quantitative real-time PCR. The spent culture media of the GC were collected for measurement of insulin growth factor 1 using ELISA.

RESULTS

The expression of BAX was significantly different after UC-MSCs-CM exposure (4.09E-7 vs. 3.74E-7, p = 0.02). No significant changes occurred in survivin, BAX/survivin ratio, and GDF9 expression after UC-MSCs-CM exposure (p 0.05). The IGF-1 level of the CM was significantly higher after the CM was used as a culture medium for GC (2.28 vs. 3.07 1.72, p 0.001). A significant positive correlation was found between survivin and GDF9 (r = 0.966, p 0.001).

CONCLUSION

IGF-1 produced by UC-MSCs-CM can work in paracrine fashion through the IGF receptor, which can inhibit BAX and maintain GDF9 production. Moreover, under the influence of UC-MSCs-CM, GC are also capable of producing IGF-1, which can impact GC through autocrine processes.

摘要

背景

脐带间充质干细胞条件培养基（UC-MSCs-CM）可产生具有抗凋亡特性的分泌组，有潜力在控制性卵巢过度刺激过程中防止颗粒细胞（GC）凋亡。

目的

观察UC-MSCs-CM对促凋亡蛋白与抗凋亡蛋白相互作用的影响以及对GC中生长分化因子9（GDF9）产生的影响。

材料与方法

从第4代脐带干细胞培养物中收集UC-MSCs-CM。培养来自23例行体外受精妇女的GC，并使其暴露于UC-MSCs-CM中24小时。然后提取GC的RNA，使用定量实时PCR分析BCL-2相关X蛋白（BAX）、生存素和GDF9的mRNA表达。收集GC的用过的培养基，用酶联免疫吸附测定法测量胰岛素生长因子1。

结果

暴露于UC-MSCs-CM后，BAX的表达有显著差异（4.09E-7对3.74E-7，p = 0.02）。暴露于UC-MSCs-CM后，生存素、BAX/生存素比值和GDF9表达无显著变化（p>0.05）。当CM用作GC的培养基后，CM中的IGF-1水平显著更高（2.28对3.07±1.72，p<0.001）。发现生存素与GDF9之间存在显著正相关（r = 0.966，p<0.001）。