Cryopreservation and recovery of human endometrial epithelial cells with high viability, purity, and functional fidelity
| Autor: | Chen, Joseph C, Hoffman, Jacquelyn R, Arora, Ripla, Perrone, Lila A, Gonzalez-Gomez, Christian J, Vo, Kim Chi, Laird, Diana J, Irwin, Juan C, Giudice, Linda C |
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| Rok vydání: | 2016 |
| Předmět: |
Cryopreservation
Cultured Cell Survival Cells Clinical Sciences Cell Polarity Epithelial Cells Cell Separation Permeability Tight Junctions Paediatrics and Reproductive Medicine stomatognathic diseases Endometrium Phenotype Gene Expression Regulation freezing medium endometrial epithelial cells endometrial epithelium Public Health and Health Services Humans Female Obstetrics & Reproductive Medicine Cell Shape Biomarkers |
| Zdroj: | Fertility and sterility, vol 105, iss 2 |
| Popis: | ObjectiveTo develop a protocol for cryopreservation and recovery of human endometrial epithelial cells (eECs) retaining molecular and functional characteristics of endometrial epithelium in vivo.DesignIn vitro study using human endometrial cells.SettingUniversity research laboratory.Patient(s)Endometrial biopsies were obtained from premenopausal women undergoing benign gynecologic procedures.Intervention(s)Primary eECs were cryopreserved in 1% fetal bovine serum/10% dimethylsulfoxide in Defined Keratinocyte Serum-Free Medium (KSFM). Recovered cells were observed for endometrial stromal fibroblast (eSF) contamination and subsequently evaluated for morphology, gene expression, and functional characteristics of freshly cultured eECs and in vivo endometrial epithelium.Main outcome measure(s)Analysis of eEC morphology and the absence of eSF contamination; evaluation of epithelial-specific gene and protein expression; assessment of epithelial polarity.Result(s)Endometrial epithelial cells recovered after cryopreservation (n = 5) displayed epithelial morphology and expressed E-cadherin (CDH1), occludin (OCLN), claudin1 (CLDN1), and keratin18 (KRT18). Compared with eSF, recovered eECs displayed increased (P |
| Databáze: | OpenAIRE |
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