Stemness maintenance of stem cells derived from human exfoliated deciduous teeth (SHED) in 3D spheroid formation through the TGF-β/Smad signaling pathway
1Longgang E.N.T. Hospital & Shenzhen Key Laboratory of E.N.T., Institute of E.N.T., 518172 Shenzhen, Guangdong, China
2Shenzhen Longgang Institute of Stomatology, 518172 Shenzhen, Guangdong, China
3Longgang District People’s Hospital of Shenzhen, 518116 Shenzhen, Guangdong, China
4Xuzhou Maternity and Child Health Care Hospital, 221009 Xuzhou, Jiangsu, China
DOI: 10.22514/jocpd.2023.081 Vol.47,Issue 6,November 2023 pp.74-85
Submitted: 26 December 2022 Accepted: 12 May 2023
Published: 03 November 2023
† These authors contributed equally.
Mesenchymal stem cells (MSCs) have shown great potential as important therapeutic tools for dental pulp tissue engineering, with the maintenance and enhancement of their stemness being crucial for successful therapeutic application in vivo and three-dimensional (3D) spheroid formation considered a reliable technique for enhancing their pluripotency. Human exfoliated deciduous tooth stem cells (SHED) were cultured in a low attachment plate to form aggregates for five days. Then, the resulting spheroids were analyzed for pluripotent marker expression, paracrine secretory function, proliferation, signaling pathways involved, and distribution of key proteins within the spheroids. The results indicated that 3D spheroid formation significantly increased the activation of the transforming growth factor beta (TGF-β)/Smad signaling pathway and upregulated the secretion and mRNA expression levels of TGF-β, which in turn enhanced the expression of pluripotency markers in SHED spheroids. The activation of the TGF-β/Smad signaling pathway through 3D spheroid formation was found to preserve the stemness properties of SHED. Thus, understanding the mechanisms behind pluripotency maintenance of SHED culture through 3D spheroid formation could have implications for the therapeutic application of MSCs in regenerative medicine and tissue engineering.
3D spheroid culture; Stemness maintenance; Stem cells derived from human exfoliated deciduous teeth (SHED); TGF-β/Smad signaling
Hongwen Li,Jing Jiang,Haiying Kong,Wenbo Wu,Xiaomin Shao,Shuqi Qiu,Xianhai Zeng,Qinghong Zhong,Xinhui Yao,Xiantao Zeng,Lingshan Gou,Jian Xu. Stemness maintenance of stem cells derived from human exfoliated deciduous teeth (SHED) in 3D spheroid formation through the TGF-β/Smad signaling pathway. Journal of Clinical Pediatric Dentistry. 2023. 47(6);74-85.
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