Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, People's Republic of China.
The First Affiliated Hospital of Huzhou University, Huzhou, Zhejiang, People's Republic of China.
Arch Gynecol Obstet. 2024 Feb;309(2):551-563. doi: 10.1007/s00404-023-07254-8. Epub 2023 Oct 23.
Although adenomyosis is a common and benign gynecological disease, the specific pathogenesis of this condition is yet to be fully elucidated. It is difficult to culture primary cells of the ectopic endometrial epithelia and stroma from human adenomyosis lesions. Most of the previous of studies on adenomyosis were based on primary eutopic endometrium cells. However, as yet, no efficient protocols have been developed for the isolation, culture or purification of primary ectopic epithelial and stromal cells from human adenomyosis lesions. Therefore, the present study aimed to develop an efficient protocol for the isolation and culture of primary ectopic epithelial and stromal cells from human adenomyosis lesions.
In the present study, we aimed to obtain ectopic endometrium tissue from human adenomyosis foci and use a simple and operable type I collagenase digestion method for primary culture. Cells were isolated by sterile cell strainer filtration and flow cytometry was performed to identify, purify, and evaluate the viability of isolated ectopic endometrial cells.
Using our method, we successfully isolated and cultured highly purified and active ectopic endometrial epithelial and stromal cells from human adenomyosis foci. Ep-CAM was expressed in ectopic epithelial cells of human adenomyosis with a purity of 93.74% and a viability of 80.58%. In addition, CD10 were robustly expressed by ectopic stromal cells in human adenomyosis. Cellular purity and viability were determined to be 96.37 and 93.49%, respectively.
Our method provides a new experimental model for studying the molecular pathogenesis of human adenomyosis.
尽管子宫腺肌病是一种常见的良性妇科疾病，但该疾病的确切发病机制尚未完全阐明。从人子宫腺肌病病变中很难培养异位子宫内膜上皮和基质的原代细胞。之前大多数关于子宫腺肌病的研究都是基于原代在位子宫内膜细胞。然而，迄今为止，尚未开发出从人子宫腺肌病病变中分离、培养或纯化原代异位上皮和基质细胞的有效方案。因此，本研究旨在开发一种从人子宫腺肌病病变中分离和培养原代异位上皮和基质细胞的有效方案。
在本研究中，我们旨在从人子宫腺肌病病灶中获得异位子宫内膜组织，并使用简单且可操作的 I 型胶原酶消化法进行原代培养。通过无菌细胞滤网分离细胞，并通过流式细胞术鉴定、纯化和评估分离的异位子宫内膜细胞的活力。
使用我们的方法，我们成功地从人子宫腺肌病病灶中分离和培养了高度纯化和活跃的异位子宫内膜上皮和基质细胞。Ep-CAM 在人子宫腺肌病的异位上皮细胞中表达，纯度为 93.74%，活力为 80.58%。此外，人子宫腺肌病的异位基质细胞中强烈表达 CD10。细胞纯度和活力分别确定为 96.37%和 93.49%。
我们的方法为研究人子宫腺肌病的分子发病机制提供了一种新的实验模型。
