Institute of Anatomy, University of Leipzig, Liebigstrasse 13, D-04103 Leipzig, Germany.
Differentiation. 2010 Apr-Jun;79(4-5):203-10. doi: 10.1016/j.diff.2010.02.004. Epub 2010 Mar 19.
Hematopoietic-like colonies develop in post-confluent granulosa cell cultures derived from bovine antral follicles. Previously, we had shown that these colonies gave rise to macrophages. In the present study, we validated the presence of somatic KIT-positive (KIT(+)) progenitor cells in colony-containing granulosa cell cultures. The cultures expressed the progenitor cell markers Sox-2, Oct 3/4, KIT, and alkaline phosphatase in western blot analysis. The successful double immunofluorescence localization of KIT and CD14, CD45, CD133, or VEGF-R2 revealed a specific subpopulation of progenitor cells. Flow cytometry showed that cells doubly positive for KIT and CD14 or CD45 comprised less than 10% of the population. The KIT(+) cells were purified by magnetic selection and differentiated with the hanging drop technique using haematopoietic differentiation medium. Pure cultures of either granulosa cells or endothelial cells were obtained. The spindle-shaped and epithelioid phenotypes indicated endothelial cell heterogeneity of microvascular source. We conclude that progenitor cells are obtained from the follicle harvest, which differentiate into endothelial cells. The cells are relevant for findings to angiogenesis and luteinization of the corpus luteum.
类造血集落可在源自牛窦卵泡的汇合后颗粒细胞培养物中发育。先前,我们已经表明这些集落可产生巨噬细胞。在本研究中,我们验证了含有集落的颗粒细胞培养物中存在体细胞 KIT 阳性(KIT(+))祖细胞。Western blot 分析表明,这些培养物表达祖细胞标志物 Sox-2、Oct 3/4、KIT 和碱性磷酸酶。KIT 和 CD14、CD45、CD133 或 VEGF-R2 的成功双重免疫荧光定位揭示了特定的祖细胞亚群。流式细胞术显示,KIT 和 CD14 或 CD45 双重阳性的细胞不到群体的 10%。KIT(+)细胞通过磁性选择进行纯化,并使用造血分化培养基通过悬滴技术进行分化。获得了纯培养的颗粒细胞或内皮细胞。纺锤形和上皮样表型表明微血管来源的内皮细胞具有异质性。我们得出结论,祖细胞是从卵泡收获中获得的,它们可分化为内皮细胞。这些细胞与血管生成和黄体黄体化有关。