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RESEARCH ARTICLE
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Harvest of functional mesenchymal stem cells derived from in vivo osteo-organoids

Shunshu Deng1,2,3 ,  Fuwei Zhu1,2,3 ,  Kai Dai1,2,3,4* ,  Jing Wang1,3,4* ,  Changsheng Liu2,3,4,5*
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1 State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China
2 Key Laboratory for Ultrafine Materials of the Ministry of Education, East China University of Science and Technology, Shanghai, China
3 Engineering Research Center for Biomedical Materials of the Ministry of Education, East China University of Science and Technology, Shanghai, China
4 Frontiers Science Center for Materiobiology and Dynamic Chemistry, East China University of Science and Technology, Shanghai, China
5 Shanghai University, Shanghai, China
BMT 2023 , 4(4), 270–279; https://doi.org/10.12336/biomatertransl.2023.04.006
Submitted: 21 September 2023 | Revised: 14 November 2023 | Accepted: 30 November 2023 | Published: 28 December 2023
Copyright © 2023 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution–NonCommercial–ShareAlike 4.0 License.
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Abstract

Bone marrow-derived mesenchymal stem cells (BM-MSCs) play a crucial role in stem cell therapy and are extensively used in regenerative medicine research. However, current methods for harvesting BM-MSCs present challenges, including a low yield of primary cells, long time of in vitro expansion, and diminished differentiation capability after passaging. Meanwhile mesenchymal stem cells (MSCs) recovered from cell banks also face issues like toxic effects of cryopreservation media. In this study, we provide a detailed protocol for the isolation and evaluation of MSCs derived from in vivo osteo-organoids, presenting an alternative to autologous MSCs. We used recombinant human bone morphogenetic protein 2-loaded gelatin sponge scaffolds to construct in vivo osteo-organoids, which were stable sources of MSCs with large quantity, high purity, and strong stemness. Compared with protocols using bone marrow, our protocol can obtain large numbers of high-purity MSCs in a shorter time (6 days vs. 12 days for obtaining passage 1 MSCs) while maintaining higher stemness. Notably, we found that the in vivo osteo-organoid-derived MSCs exhibited stronger anti-replicative senescence capacity during passage and amplification, compared to BM-MSCs. The use of osteo-organoid-derived MSCs addresses the conflict between the limitations of autologous cells and the risks associated with allogeneic sources in stem cell transplantation. Consequently, our protocol emerges as a superior alternative for both stem cell research and tissue engineering.

Keywords
anti-replicative senescence ; in vivo osteo-organoid ; mesenchymal stem cell ; recombinant human bone morphogenetic protein 2 ; stem cell therapy
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Conflict of interest
The authors declare they have no competing interests.
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