Department of Anatomy, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand.
Department of Obstetrics and Gynecology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
Hum Cell. 2021 Mar;34(2):300-309. doi: 10.1007/s13577-020-00439-2. Epub 2021 Feb 4.
Human ovarian follicular fluid (HOFF) contains proteins, extracellular matrixes necessary for growth and maturation of oocytes as well as granulosa cells. Epithelial cells and stem cells can be isolated from HOFF. However, information regarding stem cells derived from HOFF is still lacking. The objectives of the present study were to isolate, characterize, and differentiate cells derived from HOFF. HOFF was collected during the routine aspiration of oocytes in an assisted fertilization program and subjected to cell isolation, characterization, and in vitro culture. After 24 h of culture, different cell morphologies including epithelial-like-, neural-like- and fibroblast-like cells were observed. Immunocytochemistry reveals the expression of pluripotent stem cell markers (OCT4, NANOG, SSEA4), epithelial marker (CK18), FSH- and LH-receptor. For in vitro culture, the isolated cells were continuously cultured in a growth medium; alpha MEM containing 10% FBS and epidermal growth factor (EGF). After 2 weeks of in vitro culture, cells with fibroblast-like morphology dominantly grow in the culture vessels and resemble mesenchymal stem cells (MSCs). HOFF-derived cells exhibited MSC expression of CD44, CD73, CD90, CD105, CD146, and STRO-1, and were capable of differentiation into osteoblasts, chondrocytes, and adipocytes. After induction of neural differentiation, HOFF-derived cells formed spheroidal structures and expressed neural stem cell markers including Nestin, β-tubulin III, and O4. Besides, the oocyte-like structure was observed after prolonged culture of HOFF. In conclusion, cells derived from follicular fluid exhibited stem cell characteristics, which could be useful for regenerative medicine applications and cell-based therapies.
人卵巢滤泡液（HOFF）含有蛋白质、细胞外基质，这些是卵母细胞和颗粒细胞生长和成熟所必需的。上皮细胞和干细胞可从 HOFF 中分离出来。然而，关于源自 HOFF 的干细胞的信息仍然缺乏。本研究的目的是分离、鉴定和分化源自 HOFF 的细胞。HOFF 在辅助受精计划中常规抽吸卵母细胞时收集，并进行细胞分离、鉴定和体外培养。培养 24 小时后，观察到不同的细胞形态，包括上皮样、神经样和纤维母细胞样细胞。免疫细胞化学显示多能干细胞标志物（OCT4、NANOG、SSEA4）、上皮标志物（CK18）、FSH 和 LH 受体的表达。用于体外培养，分离的细胞在含有 10% FBS 和表皮生长因子（EGF）的α MEM 生长培养基中连续培养。体外培养 2 周后，以纤维母细胞样形态为主的细胞在培养容器中生长，并类似于间充质干细胞（MSCs）。HOFF 来源的细胞表现出 MSC 表达的 CD44、CD73、CD90、CD105、CD146 和 STRO-1，并能够分化为成骨细胞、软骨细胞和脂肪细胞。诱导神经分化后，HOFF 来源的细胞形成球形结构，并表达神经干细胞标志物，包括巢蛋白、β-微管蛋白 III 和 O4。此外，在 HOFF 的长时间培养后观察到卵母细胞样结构。总之，源自滤泡液的细胞表现出干细胞特征，可用于再生医学应用和基于细胞的治疗。
