title: Effects of extracellular calcium on viability and osteogenic differentiation of bone marrow stromal cells in vitro.
abstract: Bone marrow stromal cells (BMSCs) have been extensively used for tissue engineering. However, the effect of Ca(2+) on the viability and osteogenic differentiation of BMSCs has yet to be evaluated. To determine the dose-dependent effect of Ca(2+) on viability and osteogenesis of BMSCs in vitro, BMSCs were cultured in calcium-free DMEM medium supplemented with various concentrations of Ca(2+) (0, 1, 2, 3, 4, and 5 mM) from calcium citrate. Cell viability was analyzed by MTT assay and osteogenic differentiation was evaluated by alkaline phosphatase (ALP) assay, Von Kossa staining, and real-time PCR. Ca(2+) stimulated BMSCs viability in a dose-dependent manner. At slightly higher concentrations (4 and 5 mM) in the culture, Ca(2+) significantly inhibited the activity of ALP on days 7 and 14 (P < 0.01 or P < 0.05), significantly suppressed collagen synthesis (P < 0.01 or P < 0.05), and significantly elevated calcium deposition (P < 0.01) and mRNA levels of osteocalcin (P < 0.01 or P < 0.05) and osteopontin (P < 0.01 or P < 0.05). Therefore, elevated concentrations of extracellular calcium may promote cell viability and late-stage osteogenic differentiation, but may suppress early-stage osteogenic differentiation in BMSCs.
authors.affiliation: Trauma Center of the Affiliated Hospital of Hainan Medical College, 31 Long Hua Road, Haikou, 571100, China.
authors.forename: Shaowen
authors.initials: S
authors.lastname: Cheng
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Zhongqin
authors.initials: Z
authors.lastname: Lin
authors.forename: Ping
authors.initials: P
authors.lastname: Zhou
authors.forename: Xiaolei
authors.initials: X
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.forename: Qingyu
authors.initials: Q
authors.lastname: Chen
authors.forename: Dongquan
authors.initials: D
authors.lastname: Kou
authors.forename: Xiaozhou
authors.initials: X
authors.lastname: Ying
authors.forename: Yue
authors.initials: Y
authors.lastname: Shen
authors.forename: Xiaojie
authors.initials: X
authors.lastname: Cheng
authors.forename: Ziming
authors.initials: Z
authors.lastname: Yu
authors.forename: Lei
authors.initials: L
authors.lastname: Peng
authors.forename: Chuanzhu
authors.initials: C
authors.lastname: Lu
mesh_terms: D000328:Adult; D000469:Alkaline Phosphatase / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis; D003470:Culture Media; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D015675:Osteocalcin / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects*; D053495:Osteopontin / Q000378:metabolism; D017154:Stromal Cells
chemical_list: D003470:Culture Media; D015675:Osteocalcin; D053495:Osteopontin; D003094:Collagen; D000469:Alkaline Phosphatase; D002118:Calcium