Rink B Elisabeth, Amilon Karin R, Esteves Cristina L, French Hilari M, Watson Elaine, Aurich Christine, Donadeu F Xavier
Ross University School of Veterinary Medicine, Basseterre, Saint Kitts and Nevis.
The Roslin Institute, University of Edinburgh, Edinburgh, EH25 9RG, UK.
Stem Cell Res Ther. 2017 Jul 12;8(1):166. doi: 10.1186/s13287-017-0616-0.
Equine mesenchymal stromal/stem cells (MSCs) are most commonly harvested from bone marrow (BM) or adipose tissue, requiring the use of surgical procedures. By contrast, the uterus can be accessed nonsurgically, and may provide a more readily available cell source. While human endometrium is known to harbor mesenchymal precursor cells, MSCs have not been identified in equine endometrium. This study reports the isolation, culture, and characterization of MSCs from equine endometrium.
The presence of MSC and pericyte markers in endometrial sections was determined using immunohistochemistry. Stromal cells were harvested and cultured after separation of epithelial cells from endometrial fragments using Mucin-1-bound beads. For comparison, MSCs were also harvested from BM. The expression of surface markers in endometrial and BM-derived MSCs was characterized using flow cytometry and quantitative polymerase chain reaction. MSCs were differentiated in vitro into adipogenic, chondrogenic, osteogenic, and smooth muscle lineages.
Typical markers of MSCs (CD29, CD44, CD90, and CD105) and pericytes (NG2 and CD146) were localized in the equine endometrium. Both endometrial and BM MSCs grew clonally and robustly expressed MSC and pericyte markers in culture while showing greatly reduced or negligible expression of hematopoietic markers (CD45, CD34) and MHC-II. Additionally, both endometrial and BM MSCs differentiated into adipogenic, osteogenic, and chondrogenic lineages in vitro, and endometrial MSCs had a distinct ability to undergo smooth muscle differentiation.
We have demonstrated for the first time the presence of cells in equine endometrium that fulfill the definition of MSCs. The equine endometrium may provide an alternative, easily accessible source of MSCs, not only for therapeutic regeneration of the uterus, but also for other tissues where MSCs from other sources are currently being used therapeutically.
马间充质基质/干细胞（MSCs）最常从骨髓（BM）或脂肪组织中获取，这需要采用外科手术。相比之下，子宫可通过非手术方式获取，并且可能提供更容易获得的细胞来源。虽然已知人类子宫内膜含有间充质前体细胞，但尚未在马子宫内膜中鉴定出MSCs。本研究报告了从马子宫内膜中分离、培养和鉴定MSCs的过程。
使用免疫组织化学法确定子宫内膜切片中MSC和周细胞标志物的存在。使用与粘蛋白-1结合的磁珠从子宫内膜碎片中分离上皮细胞后，收获并培养基质细胞。为作比较，也从骨髓中获取MSCs。使用流式细胞术和定量聚合酶链反应来鉴定子宫内膜和骨髓来源的MSCs表面标志物的表达。将MSCs在体外分化为脂肪生成、软骨生成、成骨和平滑肌谱系。
MSCs（CD29、CD44、CD90和CD105）和周细胞（NG2和CD146）的典型标志物定位于马子宫内膜中。子宫内膜和骨髓来源的MSCs均呈克隆生长，并且在培养中强烈表达MSC和周细胞标志物，而造血标志物（CD45、CD34）和MHC-II的表达大大降低或可忽略不计。此外，子宫内膜和骨髓来源的MSCs在体外均分化为脂肪生成、成骨和软骨生成谱系，并且子宫内膜来源的MSCs具有独特的平滑肌分化能力。
我们首次证明马子宫内膜中存在符合MSCs定义的细胞。马子宫内膜可能提供一种替代的、易于获取的MSCs来源，不仅可用于子宫的治疗性再生，还可用于目前正在使用其他来源的MSCs进行治疗的其他组织。
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