Experimental Study
Effect of excess iron culturing on the proliferation and osteogenic differentiation of rat bone mesenchymal stem cells
Chen Yu, Fan Yang, Jian Li, Bin Chen, Ye Yuan, Yan Gao, Yong Li, Beibei Fei, Youjia Xu, Jiachen Peng
Published 2018-04-08
Cite as Chin J Exp Surg, 2018, 35(4): 719-722. DOI: 10.3760/cma.j.issn.1001-9030.2018.04.037
Abstract
ObjectiveTo investigate the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in the culture with excess iron.
MethodsFour-week old female Sprague-Dawley (SD) rats were used for BMSCs isolation from bone marrow. BMSCs were identified by flow cytometry and exposed to ammonium iron citrate (FAC) (50, 100 and 200 μmol/L) under osteogenic medium for 1 to 2 weeks. The proliferation and differentiation of BMSCs were analyzed by cell counting kit-8 (CCK-8), Alizarin Red S staining and real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR) tests.
ResultsThe proliferation activity of BMSCs decreased with the increase of FAC concentration. The formation of calcium nodules decreased with the increase of FAC concentration (P=0.000). The expression of osteogenic differentiation genes runt related transcription factor-2 (Runx2) decreased with the increase of FAC concentration (sham group: 1.000±0.229; 100 μmol/L FAC group: 0.198±0.044; 200 μmol/L FAC group: 0.149±0.009, P=0.001). The mRNA expression of ALP decreased with the increase of FAC concentration (sham group: 0.980±0.063; 50 μmol/L FAC group: 0.018±0.001; 100 μmol/L FAC group: 0.014±0.002; 200 μmol/L FAC group: 0.008±0.001, P=0.001). The mRNA expression of OPN decreased with the increase of FAC concentration (sham group: 0.999±0.001; 50 μmol/L FAC group: 0.170±0.072; 100 μmol/L FAC group: 0.142±0.008; 200 μmol/L FAC group: 0.117±0.028, P=0.003).
ConclusionThe ammonium iron citrate significantly inhibit the proliferation of rat BMSCs, and simultaneously suppress osteogenic differentiation of rat BMSCs dose-dependently.
Key words:
Bone marrow mesenchymal stem cells; Iron citrate; Osteogenic differentiation; Rat
Contributor Information
Chen Yu
Department of Orthopaedics, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China
Fan Yang
Department of Orthopaedics, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China
Jian Li
Center for Translational Medicine Research and Development, Shenzhen Institutes of Advanced Technology, Chinese Academy of Science, Shenzhen 518055, China
Bin Chen
Department of Orthopaedics, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China
Ye Yuan
Department of Orthopaedics, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China
Yan Gao
Department of Orthopaedics, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China
Yong Li
Department of Radiology, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China
Beibei Fei
Department of Obstetrics and Gynecology, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China
Youjia Xu
Department of Orthopaedics, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China
Jiachen Peng
Department of Joint Surgery, the Affiliated Hospital of Zunyi Medical College, Zunyi 563003, China
Department of Orthopaedics, the Second Affiliated Hospital of Soochow University, Suzhou 215000, China
