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Updated: Jun 6, 2025
Isolation and Cultivation of Mandibular Bone Marrow Mesenchymal Stem Cells in Rats
Published on: August 25, 2020
Establishment of immortalized rabbit bone marrow mesenchymal stem cells and a preliminary study of their osteogenic
Yao Zhang1, Chang Xu1, Yun Huang1
1Laboratory of Animal Center, Chongqing Medical University, Chongqing, China.
Background:
A stable and standardized source of mesenchymal stem cells is a prerequisite for bone repair tissue engineering research and application. We aimed to establish a stable cell line of bone marrow mesenchymal stem cells from New Zealand rabbits and explore their osteogenic differentiation capacity.
Methods:
Primary rabbit bone marrow mesenchymal stem cells (RBMSCs) were isolated and immortalized via retroviral expression of SV40 Large T antigen (LTA). To assess the osteogenic differentiation capacity of the cells in vitro, we studied the alkaline phosphatase (ALP) expression level and calcium deposition in bone morphogenetic protein 9 (BMP9)-induced immortalized cells using ALP staining and quantification, as well as alizarin red staining. Ectopic bone formation by the cells was assessed using micro-computed tomography (μCT) and histological examination.
Results:
The immortalized cell line we established using SV40 LTA, which we termed iRBMSCs, was non-tumorigenic and maintained long-term proliferative activity. We further discovered that BMP9 (MOI = 30) effectively induced the osteogenic differentiation capacity of iRBMSCs in vitro, and there was a synergy with GelMA hydrogel in inducing osteogenic differentiation of the iRBMSCs in vivo.
Conclusion:
We confirmed that iRBMSCs are promising as a stable cell line source for bone defect repair engineering.
