Differences in metabolism, proliferation, differentiation of adipose-derived mesenchymal stem cells, and differentiation into vascular smooth muscle cells between male and female rats

doi:10.12307/2026.060

Chinese Journal of Tissue Engineering Research ›› 2026, Vol. 30 ›› Issue (7): 1687-1698.doi: 10.12307/2026.060

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Differences in metabolism, proliferation, differentiation of adipose-derived mesenchymal stem cells, and differentiation into vascular smooth muscle cells between male and female rats

Wang Qiuhua1, Du Ziwei1, Wang Wenshuang2, Zhao Dongmei1, Zhang Xiaoqing1

1School of Basic Medical Sciences, Binzhou Medical University, Department of Human Anatomy and Histoembryology, Joint Laboratory for International Cooperation in Biomaterials and Tissue Regeneration, Yantai 264003, Shandong Province, China; 2Department of Gynecology, Yantai Yuhuanding Hospital, Yantai 264000, Shandong Province, China

Received:2025-01-15 Revised:2025-05-09 Accepted:2025-05-29 Online:2026-03-08 Published:2025-08-19
Contact: Zhang Xiaoqing, MD, Professor, School of Basic Medical Sciences, Binzhou Medical University, Department of Human Anatomy and Histoembryology, Joint Laboratory for International Cooperation in Biomaterials and Tissue Regeneration, Yantai 264003, Shandong Province, China; Zhao Dongmei, MD, Professor, School of Basic Medical Sciences, Binzhou Medical University, Department of Human Anatomy and Histoembryology, Joint Laboratory for International Cooperation in Biomaterials and Tissue Regeneration, Yantai 264003, Shandong Province, China
About author:Wang Qiuhua, MS, School of Basic Medical Sciences, Binzhou Medical University, Department of Human Anatomy and Histoembryology, Joint Laboratory for International Cooperation in Biomaterials and Tissue Regeneration, Yantai 264003, Shandong Province, China; Du Ziwei, MS, School of Basic Medical Sciences, Binzhou Medical University, Department of Human Anatomy and Histoembryology, Joint Laboratory for International Cooperation in Biomaterials and Tissue Regeneration, Yantai 264003, Shandong Province, China Wang Qiuhua and Du Ziwei contributed equally to this article.
Supported by:
Taishan Scholar Youth Project, No. tsqn202103111 (to ZXQ); Shandong Natural Science Foundation Youth Fund, No. ZR2021QC034 (to ZXQ); Yantai Province-level Talent Matching Fund, No. 1007-10073802 (to ZXQ); Shandong Overseas High-level Talent Workstation Fund, No. 2023119005 (to ZXQ)

Abstract

Abstract: BACKGROUND: In the structure of the vascular wall, vascular smooth muscle cells maintain pressure stability by regulating vascular tension, ensuring blood supply to tissues. Vascular smooth muscle cells exhibit high plasticity and are a critical cell type for vascular regeneration. However, mature vascular smooth muscle cells are difficult to obtain and have limited expansion capacity in vitro. In contrast, adipose-derived mesenchymal stem cells are easily accessible, possess strong expansion capabilities, and have the potential to differentiate into vascular smooth muscle cells. Currently, whether there are differences in biological characteristics such as metabolic activity, proliferation, colony formation, and the ability to differentiate into vascular smooth muscle cells between adipose-derived mesenchymal stem cells derived from females and males remains to be thoroughly investigated.
OBJECTIVE: To compare the differences in biological characteristics of adipose-derived mesenchymal stem cells from female and male rats in metabolic activity, proliferation, differentiation ability, and differentiation potential into vascular smooth muscle cells.
METHODS: (1) Adipose tissue was harvested from the inguinal region of both female and male rats, and adipose-derived mesenchymal stem cells were isolated using collagenase digestion. Surface markers (CD90, CD29, and CD45) were detected by flow cytometry. The biological characteristics of adipose-derived mesenchymal stem cells from both sexes were compared using WST-1 assay, cell doubling assay, colony formation assay, and adipogenic, osteogenic, and chondrogenic differentiation assays. (2) Adipose-derived mesenchymal stem cells from female and male rats were induced to differentiate into vascular smooth muscle cells using 5 ng/mL transforming growth factor-β. The expression of α-SMA, SM22α, Calponin, Caldesmon, SMMHC, and Smoothelin mRNAs was detected by qRT-PCR, while the expression of α-SMA, Caldesmon, Smoothelin proteins was assessed by immunofluorescence staining. The contractile function of vascular smooth muscle cells was evaluated using a collagen gel contraction assay.
RESULTS AND CONCLUSION: (1) After 3 days of primary culture, microscopic observation revealed that adipose-derived stem cells from both female and male rats exhibited a uniform, spindle-shaped morphology. (2) Flow cytometry analysis showed that rat adipose-derived mesenchymal stem cells highly expressed CD90 and CD29, while CD45 expression was low. (3) Compared to adipose-derived mesenchymal stem cells from male rats, female rat adipose-derived mesenchymal stem cells demonstrated significantly higher metabolic activity (P < 0.05). (4) Adipose-derived mesenchymal stem cells from both sexes showed similar doubling time, colony formation ability, and adipogenic, osteogenic, and chondrogenic differentiation potential. (5) After induction of differentiation into vascular smooth muscle cell, at the gene level, female rat adipose-derived mesenchymal stem cells exhibited significantly higher expression of α-SMA, Caldesmon, and SMMHC compared to male rat adipose-derived mesenchymal stem cells (P < 0.05). At the protein level, female rat adipose-derived mesenchymal stem cells showed significantly higher expression of α-SMA (P < 0.05). However, in terms of contractile function, adipose-derived mesenchymal stem cells from both sexes displayed similar performance. In conclusion, female rat adipose-derived mesenchymal stem cells demonstrated superior metabolic activity and a greater potential for differentiation into vascular smooth muscle cells compared to male rat adipose-derived mesenchymal stem cells.

Key words: adipose-derived mesenchymal stem cell, vascular smooth muscle cell, induced differentiation, female, male, small diameter artificial blood vessel

CLC Number:

Wang Qiuhua, Du Ziwei, Wang Wenshuang, Zhao Dongmei, Zhang Xiaoqing. Differences in metabolism, proliferation, differentiation of adipose-derived mesenchymal stem cells, and differentiation into vascular smooth muscle cells between male and female rats [J]. Chinese Journal of Tissue Engineering Research, 2026, 30(7): 1687-1698.

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Differences in metabolism, proliferation, differentiation of adipose-derived mesenchymal stem cells, and differentiation into vascular smooth muscle cells between male and female rats

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