Low-serum culture with novel medium promotes maxillary/mandibular bone marrow stromal cell proliferation and osteogenic differentiation ability
Low-serum culture with novel medium promotes maxillary/mandibular bone marrow stromal cell proliferation and osteogenic differentiation ability
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新型培养基低血清培养促进上颌/下颌骨髓基质细胞增殖和成骨分化能力
DOI：
10.1007/s00784-017-2073-7
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Nishimura M
中科院分区：
文献类型：
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作者：
Suehiro F;Ishii M;Asahina I;Murata H;Nishimura M
本研究旨在探讨低血清STK 2培养液对人上颌/下颌骨骨髓基质细胞（MBMSCs）分离及向成骨细胞分化的影响。这些细胞在无血清培养基或含1%胎牛血清的STK 2培养基（低血清）或含10%胎牛血清的α-MEM（对照）中培养。对培养表面上的增殖、细胞表面抗原表达以及神经嵴和成骨标志物的mRNA水平进行了检查。碱性磷酸酶法和茜素红染色法检测成骨细胞分化能力。结果低血清组和对照组骨髓间充质干细胞表达CD 73、CD 90和CD 105，表达CD 14、CD 34、CD 45、CD 271和HLA-DR。低血清组的骨髓间充质干细胞增殖率高于对照组。诱导成骨分化后，低血清MBMSCs碱性磷酸酶活性和骨钙素mRNA水平高于对照组，茜素红染色强于对照组。结论低血清预培养的MBMSC较对照组具有更高的细胞增殖和成骨分化能力，提示低血清STK 2培养可能有助于促进MBMSC的增殖和成骨分化。这种方法比传统方法需要更少的自体血液收集用于细胞扩增，从而减轻了患者的负担。
ObjectivesThe purpose of this study was to evaluate the effect of low-serum STK2 medium on the isolation and osteogenic differentiation of human maxillary/mandibular bone marrow stromal cells (MBMSCs).Materials and methodsHuman MBMSCs were obtained from patients undergoing dental implant treatment. These cells were cultured in serum-free medium or STK2 medium containing 1 % fetal bovine serum (low-serum) or α-MEM containing 10 % fetal bovine serum (control). Proliferation on the culture surface, cell surface antigen expression, and mRNA levels of neural crest and osteogenic markers were examined. Alkaline phosphatase assay and Alizarin red staining were used to assess osteogenic differentiation potential. Immunoblotting analysis was performed to detect ERK phosphorylation.ResultsLow-serum and control MBMSCs were positive for CD73, CD90, and CD105 and negative for CD14, CD34, CD45, CD271, and HLA-DR. CD140a was absent in low-serum cells but present in control cells. Low-serum MBMSCs proliferated more than control MBMSCs. After induction of osteogenic differentiation, alkaline phosphatase activity and osteocalcin mRNA levels were higher in low-serum MBMSCs than in control cells, and Alizarin red staining was stronger in low-serum MBMSCs than in control cells. Low-serum culture promoted ERK phosphorylation.ConclusionsMBMSCs precultured in low-serum medium exhibited a greater cumulative cell number and a higher osteogenic differentiation capacity than those cultured in control medium.Clinical relevanceThese findings indicate that low-serum STK2 culture might be useful to promote MBMSC proliferation and osteogenic differentiation. This method requires less autologous blood collection for cell expansion than conventional methods, thus reducing the burden on patients.