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Differentiation of human endometrial stem cells encapsulated in alginate hydrogel into oocyte-like cells

Ghasemi, Diba, Ebrahimi-Barough, Somayeh, Nekoofar, Mohammad Hossein, Mohamadnia, Abdolreza, Lotfibakhshaiesh, Nasrin, Bahrami, Naghmeh, Karimi, Roya, Nooshabadi, Vajihe Taghdiri, Azami, Mahmoud, Hasanzadeh, Elham and Ai, Jafar 2023. Differentiation of human endometrial stem cells encapsulated in alginate hydrogel into oocyte-like cells. BioImpacts 13 (3) , pp. 229-240. 10.34172/bi.2022.23960

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Abstract

Introduction: Human endometrial mesenchymal stem cells (hEnMSCs) are a rich source of mesenchymal stem cells (MSCs) with multi-lineage differentiation potential, making them an intriguing tool in regenerative medicine, particularly for the treatment of reproductive and infertility issues. The specific process of germline cell-derived stem cell differentiation remains unknown, the aim is to study novel ways to achieve an effective differentiation method that produces adequate and functioning human gamete cells. Methods: We adjusted the optimum retinoic acid (RA) concentration for enhancement of germ cell-derived hEnSCs generation in 2D cell culture after 7 days in this study. Subsequently, we developed a suitable oocyte-like cell induction media including RA and bone morphogenetic protein 4 (BMP4), and studied their effects on oocyte-like cell differentiation in 2D and 3D cell culture media utilizing cells encapsulated in alginate hydrogel. Results: Our results from microscopy analysis, real-time PCR, and immunofluorescence tests revealed that 10 µM RA concentration was the optimal dose for inducing germ-like cells after 7 days. We examined the alginate hydrogel structural characteristics and integrity by rheology analysis and SEM microscope. We also demonstrated encapsulated cell viability and adhesion in the manufactured hydrogel. We propose that in 3D cell cultures in alginate hydrogel, an induction medium containing 10 µM RA and 50 ng/mL BMP4 can enhance hEnSC differentiation into oocyte-like cells. Conclusion: The production of oocyte-like cells using 3D alginate hydrogel may be viable in vitro approach for replacing gonad tissues and cells.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Dentistry
Publisher: Tabriz University of Medical Sciences
ISSN: 2228-5660
Date of First Compliant Deposit: 20 March 2023
Date of Acceptance: 4 December 2021
Last Modified: 02 Aug 2023 16:33
URI: https://orca.cardiff.ac.uk/id/eprint/157820

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