Department of Obstetrics and Gynaecology, University Medical Centre Ljubljana, Šlajmerjeva 3, 1000 Ljubljana, Slovenia.
Department of Molecular Diagnostics, Institute of Oncology Ljubljana, Zaloška 2, 1000 Ljubljana, Slovenia.
Biomed Res Int. 2014;2014:508216. doi: 10.1155/2014/508216. Epub 2014 Mar 3.
After removal of oocytes for in vitro fertilization, follicular aspirates which are rich in somatic follicular cells are discarded in daily medical practice. However, there is some evidence that less differentiated cells with stem cell characteristics are present among aspirated follicular cells (AFCs). The aim of this study was to culture AFCs in vitro and to analyze their gene expression profile. Using the RT2 Profiler PCR array, we investigated the expression profile of 84 genes related to stemness, mesenchymal stem cells (MCSs), and cell differentiation in AFCs enriched by hypoosmotic protocol from follicular aspirates of infertile women involved in assisted reproduction programme in comparison with bone marrow-derived mesenchymal stem cells (BM-MSCs) and fibroblasts. Altogether the expression of 57 genes was detected in AFCs: 16 genes (OCT4, CD49f, CD106, CD146, CD45, CD54, IL10, IL1B, TNF, VEGF, VWF, HDAC1, MITF, RUNX2, PPARG, and PCAF) were upregulated and 20 genes (FGF2, CASP3, CD105, CD13, CD340, CD73, CD90, KDR, PDGFRB, BDNF, COL1A1, IL6, MMP2, NES, NUDT6, BMP6, SMURF2, BMP4, GDF5, and JAG1) were downregulated in AFCs when compared with BM-MSCs. The genes which were upregulated in AFCs were mostly related to MSCs and connected with ovarian function, and differed from those in fibroblasts. The cultured AFCs with predominating granulosa cells were successfully in vitro differentiated into adipogenic-, osteogenic-, and pancreatic-like cells. The upregulation of some MSC-specific genes and in vitro differentiation into other types of cells indicated a subpopulation of AFCs with specific stemness, which was not similar to those of BM-MSCs or fibroblasts.
在体外受精取出卵子后,富含体腔卵泡细胞的卵泡抽吸物在日常医疗实践中被丢弃。然而,有一些证据表明,在抽吸的卵泡细胞(AFC)中存在分化程度较低、具有干细胞特征的细胞。本研究的目的是在体外培养 AFC,并分析其基因表达谱。我们使用 RT2 Profiler PCR 阵列,比较了从参与辅助生殖计划的不孕妇女的卵泡抽吸物中通过低渗方案富集的 AFC 与骨髓间充质干细胞(BM-MSCs)和成纤维细胞之间与干性、间充质干细胞(MCSs)和细胞分化相关的 84 个基因的表达谱。总共在 AFC 中检测到 57 个基因的表达:16 个基因(OCT4、CD49f、CD106、CD146、CD45、CD54、IL10、IL1B、TNF、VEGF、VWF、HDAC1、MITF、RUNX2、PPARG 和 PCAF)上调,20 个基因(FGF2、CASP3、CD105、CD13、CD340、CD73、CD90、KDR、PDGFRB、BDNF、COL1A1、IL6、MMP2、NES、NUDT6、BMP6、SMURF2、BMP4、GDF5 和 JAG1)下调与 BM-MSCs 相比,在 AFC 中。在 AFC 中上调的基因主要与 MSC 相关,并与卵巢功能有关,与成纤维细胞中的基因不同。培养的以颗粒细胞为主的 AFC 可成功体外分化为脂肪细胞、成骨细胞和胰腺样细胞。一些 MSC 特异性基因的上调和向其他类型细胞的体外分化表明,AFC 存在具有特定干性的亚群,与 BM-MSCs 或成纤维细胞不同。