Culture of human ovarian tissue in xeno-free conditions using laminin components of the human ovarian extracellular matrix.

Journal of Assisted Reproduction and Genetics
J HaoP Damdimopoulou

Abstract

Our purpose was to identify human ovarian extracellular matrix (ECM) components that would support in vitro culture of human ovarian tissue and be compatible with possible future clinical applications. We characterized ovarian expression of laminins and selected three laminin tripeptides for culture experiments to be compared with Matrigel, an undefined and animal-based mixture of ECM components. Expression of the 12 laminin genes was determined on transcript and protein levels using cortical tissue samples (n = 6), commercial ovary RNA (n = 1), follicular fluid granulosa cells (n = 20), and single-cell RNA-sequencing data. Laminin 221 (LN221), LN521, LN511, and their mixture were chosen for a 7-day culture experiment along with Matrigel using tissue from 17 patients. At the end of the culture, follicles were evaluated by scoring and counting from serial tissue sections, apoptosis measured using in situ TUNEL assay, proliferation by Ki67 staining, and endocrine function by quantifying steroids in culture media using UPLC-MS/MS. Approximately half of the cells in ovarian cortex expressed at least one laminin gene. The overall most expressed laminin α-chains were LAMA2 and LAMA5, β-chains LAMB1 and LAMB2, and γ-chain LAMC1. In cu...Continue Reading

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Citations

Jan 11, 2021·European Journal of Obstetrics, Gynecology, and Reproductive Biology·Zeinab GhezelayaghBita Ebrahimi
Sep 14, 2021·Pathology Oncology Research : POR·Bowen Diao, Ping Yang

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