Department of Reproductive Medicine and Gynaecology, University Medical Centre Utrecht, Heidelberglaan 100, CX Utrecht 3584, the Netherlands.
Department of Development and Regeneration, Cluster of Stem Cell and Developmental Biology, Unit of Stem Cell Research, KU Leuven (University of Leuven), Herestraat 49, Leuven 3001, Belgium.
Reprod Biomed Online. 2020 Sep;41(3):465-473. doi: 10.1016/j.rbmo.2020.03.019. Epub 2020 Apr 29.
Can organoids be established from endometrial tissue of infertile women and does tissue cryopreservation allow for establishment of organoids comparable to organoids derived from freshly biopsied endometrial tissue?
Endometrial tissue was obtained from six infertile women through minimally invasive biopsy using a Pipelle catheter and subjected to organoid development, immediately after biopsy as well as after tissue cryopreservation. Organoid formation efficiency, morphology, expandability potential, endometrial marker expression (immunostaining and reverse transcription quantitative real-time polymerase chain reaction) and hormonal responsiveness (after oestradiol and progesterone treatment) were assessed.
Organoids established from both fresh and frozen tissue at comparable efficiency could be passaged long-term and showed similar morphology, i.e. cystic with a central lumen lined by a single epithelial cell layer. They also exhibited comparable expression of endometrial markers and proliferative activity (Ki67 expression). Finally, organoids from freshly biopsied and cryopreserved endometrial tissue showed similar responses to oestradiol and progesterone treatment.
Organoids can be established from cryopreserved endometrial tissue of infertile women and cryopreservation of the biopsy does not affect organoid formation and overall organoid characteristics. Cryopreservation of biopsies for later organoid development facilitates sample collection from any fertility clinic, not just the ones near an organoid laboratory.
能否从不孕女性的子宫内膜组织中建立类器官,组织冷冻保存是否允许建立与从新鲜活检的子宫内膜组织中获得的类器官相当的类器官?
通过使用 Pipelle 导管对六名不孕女性进行微创活检获得子宫内膜组织,并在活检后立即以及组织冷冻保存后进行类器官发育。评估类器官形成效率、形态、扩展性潜力、子宫内膜标志物表达(免疫染色和逆转录定量实时聚合酶链反应)和激素反应性(雌激素和孕激素处理后)。
从新鲜和冷冻组织中以相当的效率建立的类器官可以长期传代,并且具有相似的形态,即囊性,中央有一个由单层上皮细胞衬里的腔。它们还表现出类似的子宫内膜标志物表达和增殖活性(Ki67 表达)。最后,新鲜活检和冷冻保存的子宫内膜组织中的类器官对雌激素和孕激素处理的反应相似。
可以从不孕女性的冷冻保存的子宫内膜组织中建立类器官,并且活检的冷冻保存不会影响类器官的形成和整体类器官特征。冷冻保存活检标本以进行以后的类器官发育,便于从任何生育诊所收集样本,而不仅仅是靠近类器官实验室的诊所。