Co-culture of STRO1 + human gingival mesenchymal stem cells and human umbilical vein endothelial cells in 3D spheroids: enhanced in vitro osteogenic and angiogenic capacities

Yushan Liu; Pei Chen; Tengfei Zhou; Jincheng Zeng; Ziyi Liu; Ruijie Wang; Yiwei Xu; Wuwei Yin; Mingdeng Rong

doi:10.3389/fcell.2024.1378035

Co-culture of STRO1 + human gingival mesenchymal stem cells and human umbilical vein endothelial cells in 3D spheroids: enhanced in vitro osteogenic and angiogenic capacities

Front Cell Dev Biol. 2024 May 6:12:1378035. doi: 10.3389/fcell.2024.1378035. eCollection 2024.

Authors

Yushan Liu^#¹, Pei Chen^#¹, Tengfei Zhou¹, Jincheng Zeng², Ziyi Liu¹, Ruijie Wang¹, Yiwei Xu³, Wuwei Yin¹, Mingdeng Rong¹

Affiliations

¹ Department of Periodontology and Implantology, Stomatological Hospital, School of Stomatology, Southern Medical University, Guangzhou, China.
² Guangdong Provincial Key Laboratory of Medical Molecular Diagnostics, Guangdong Key Laboratory of Medical Bioactive Molecular Developmental and Translational Research, Guangdong Medical University, Dongguan, China.
³ Guangzhou Regenerative Medicine and Health Guangdong Laboratory, CAS Key Laboratory of Regenerative Biology, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Science, Guangzhou, China.

^# Contributed equally.

Abstract

Stem cell spheroid is a promising graft substitute for bone tissue engineering. Spheroids obtained by 3D culture of STRO1+ Gingival Mesenchymal Stem Cells (sGMSCs) (sGMSC spheroids, GS) seldom express angiogenic factors, limiting their angiogenic differentiation in vivo. This study introduced a novel stem cell spheroid with osteogenic and angiogenic potential through 3D co-culture of sGMSCs and Human Umbilical Vein Endothelial Cells (HUVECs) (sGMSC/HUVEC spheroids, GHS). GHS with varying seeding ratios of sGMSCs to HUVECs (GHR) were developed. Cell fusion within the GHS system was observed via immunofluorescence. Calcein-AM/PI staining and chemiluminescence assay indicated cellular viability within the GHS. Furthermore, osteogenic and angiogenic markers, including ALP, OCN, RUNX2, CD31, and VEGFA, were quantified and compared with the control group comprising solely of sGMSCs (GS). Incorporating HUVECs into GHS extended cell viability and stability, initiated the expression of angiogenic factors CD31 and VEGFA, and upregulated the expression of osteogenic factors ALP, OCN, and RUNX2, especially when GHS with a GHR of 1:1. Taken together, GHS, derived from the 3D co-culture of sGMSCs and HUVECs, enhanced osteogenic and angiogenic capacities in vitro, extending the application of cell therapy in bone tissue engineering.

Keywords: Gingival mesenchymal stem cells; STRO1; angiogenic differentiation; human umbilical Vein endothelial cells; osteogenic differentiation; stem cell spheroids; three-dimensional culture.

Grants and funding

The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. This work was supported by the Guangdong International Scientific Research Cooperation Project, Grant/Award Number 2022A0505050037, the Guangzhou Basic and Applied Basic Research Foundation, Grant/Award Number 2023A04J0429 and the Scientific Research Project of Southern Medical University Stomatological Hospital, Grant/Award Number PY2022029.