Isolation and Culture of Epithelial Progenitors and Mesenchymal Stem Cells from Human Endometrium
Isolation and Culture of Epithelial Progenitors and Mesenchymal Stem Cells from Human Endometrium
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DOI：
10.1095/biolreprod.108.075226
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发表时间：
2009-06-01
影响因子：
3.6
通讯作者：
Wu, Di
中科院分区：
文献类型：
--
作者：
Gargett, Caroline E.;Schwab, Kjiana E.;Wu, Di
人类子宫内膜是一种高度再生的组织，在女性的生殖年龄期间经历400多个生长、分化和脱落周期。子宫内膜再生可能是由成体干/祖细胞介导的。本研究探讨了从人子宫内膜获得的单个克隆形成上皮细胞和基质细胞的关键干细胞特性。从15名月经周期正常的妇女子宫切除组织中获得子宫内膜上皮或基质细胞的单细胞悬液，并以克隆密度（10个细胞/cm（2））或有限稀释培养。成体干细胞的自我更新，高增殖能力，和分化的单个上皮细胞和基质细胞进行了评估收获单个集落和进行系列克隆培养，连续传代，并在分化诱导培养基中培养，分别。采用RT-PCR、免疫细胞化学和流式细胞术检测细胞系分化标志物。罕见的单个人子宫内膜EpCAM（+）上皮细胞和EpCAM（-）基质细胞通过连续克隆> 3次显示自我更新，并在培养中经历了4个月以上的> 30次群体倍增。克隆来源的上皮细胞在三维培养中分化为细胞角蛋白（+）腺样结构。单个基质细胞是多能的，因为它们的后代分化成平滑肌细胞、脂肪细胞、软骨细胞和成骨细胞。基质克隆表达间充质干细胞（MSC）标志物ITGB 1（CD 29）、CD 44、NT 5E（CD 73）、THY 1（CD 90）、ENG（CD 105）、PDGFRB（CD 140 B）、MCAM（CD 146），但不表达内皮或造血标志物PECAM 1（CD 31）、CD 34、PTPRC（CD 45）。成人子宫内膜含有罕见的上皮祖细胞和MSC，可能是其巨大再生能力的原因，这也可能在子宫内膜异位症和子宫内膜癌的发展中起关键作用。人子宫内膜可以为基于细胞的疗法提供现成的MSC来源。
Human endometrium is a highly regenerative tissue undergoing more than 400 cycles of growth, differentiation, and shedding during a woman's reproductive years. Endometrial regeneration is likely mediated by adult stem/progenitor cells. This study investigated key stem cell properties of individual clonogenic epithelial and stromal cells obtained from human endometrium. Single-cell suspensions of endometrial epithelial or stromal cells were obtained from hysterectomy tissues from 15 women experiencing normal menstrual cycles, and were cultured at clonal density ( 10 cells/cm(2)) or limiting dilution. The adult stem cell properties-self-renewal, high proliferative potential, and differentiation of single epithelial and stromal cells-were assessed by harvesting individual colonies and undertaking serial clonal culture, serial passaging, and culture in differentiation-induction media, respectively. Lineage differentiation markers were examined by RT-PCR, immunocytochemistry, and flow cytometry. Rare single human endometrial EpCAM(+) epithelial cells and EpCAM(-) stromal cells demonstrated self-renewal by serially cloning > 3 times and underwent > 30 population doublings over 4 mo in culture. Clonally derived epithelial cells differentiated into cytokeratin(+) gland-like structures in three dimensional culture. Single stromal cells were multipotent, as their progeny differentiated into smooth muscle cells, adipocytes, chondrocytes, and osteoblasts. Stromal clones expressed mesenchymal stem cell (MSC) markers ITGB1 (CD29), CD44, NT5E (CD73), THY1 (CD90), ENG (CD105), PDGFRB (CD140B), MCAM (CD146) but not endothelial or hemopoietic markers PECAM1 (CD31), CD34, PTPRC (CD45). Adult human endometrium contains rare epithelial progenitors and MSCs, likely responsible for its immense regenerative capacity, which may also have critical roles in the development of endometriosis and endometrial cancer. Human endometrium may provide a readily available source of MSCs for cell-based therapies.