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在自体血清中培养的人骨髓间充质基质细胞的成骨潜能:一项初步研究。

Osteogenic potential of human bone marrow-derived mesenchymal stromal cells cultured in autologous serum: a preliminary study.

作者信息

Takeda Akira, Yamazaki Yasuharu, Baba Kyoko, Ishiguro Masashi, Aoyagi Kazuya, Ikemoto Shigehiro, Uchinuma Eiju

机构信息

Department of Plastic and Aesthetic Surgery, School of Medicine, Kitasato University, Kanagawa, Japan.

出版信息

J Oral Maxillofac Surg. 2012 Aug;70(8):e469-76. doi: 10.1016/j.joms.2012.03.021.

Abstract

PURPOSE

As part of the authors' research on potential osteogenesis by filling bone defects with human bone marrow-derived mesenchymal stromal cells (hBM-MSCs) in patients with cleft lip and palate, they examined the cytoproliferative potential and cytobiological activity of hBM-MSCs in vitro and their osteogenic potential in vivo without performing osteoinduction.

MATERIALS AND METHODS

The hBM-MSCs were collected from iliac cancellous bone and then used in primary culture, followed by 2 subcultures using an autologous serum (AS)-containing medium and a fetal bovine serum (FBS)-containing medium. Cytoproliferative potential and cytobiological activity as expressed by bone markers (alkaline phosphatase and osteocalcin) in hBM-MSCs cultured in the AS-containing medium (AS-cultured hBM-MSCs) and the FBS-containing medium (FBS-cultured hBM-MSCs) were examined in vitro, and the osteogenic potential of AS- and FBS-cultured hBM-MSCs was examined in mice.

RESULTS

On day 6 of the second subculture, the number of hBM-MSCs per milliliter of specimen from 8 pediatric patients was significantly larger (P < .05) in FBS-cultured compared with AS-cultured hBM-MSCs. The alkaline phosphatase activity of hBM-MSCs was significantly greater (P < .05) when cultured in the AS-containing medium compared with the FBS-containing medium. The in vivo study showed the formation of an osteoid-like matrix rather than definite bone tissue.

CONCLUSIONS

  1. FBS is appropriate for the cytoproliferation of hBM-MSCs; 2) the AS-containing medium is likely to have a good possibility of inducing the differentiation of hBM-MSCs; and 3) AS-cultured hBM-MSCs contain a group of cells that spontaneously differentiate into an osteoid-like matrix without performing osteoinduction.
摘要

目的

作为作者关于在唇腭裂患者中用人骨髓间充质基质细胞(hBM-MSCs)填充骨缺损时潜在成骨作用研究的一部分,他们在未进行骨诱导的情况下,检测了hBM-MSCs在体外的细胞增殖潜能和细胞生物学活性以及在体内的成骨潜能。

材料与方法

从髂骨松质骨采集hBM-MSCs,然后进行原代培养,接着使用含自体血清(AS)的培养基和含胎牛血清(FBS)的培养基进行2次传代培养。检测在含AS培养基(AS培养的hBM-MSCs)和含FBS培养基(FBS培养的hBM-MSCs)中培养的hBM-MSCs的细胞增殖潜能和由骨标志物(碱性磷酸酶和骨钙素)表达的细胞生物学活性,并在小鼠体内检测AS培养和FBS培养的hBM-MSCs的成骨潜能。

结果

在第二次传代培养的第6天,与AS培养的hBM-MSCs相比,8例儿科患者每毫升标本中FBS培养的hBM-MSCs数量显著更多(P < 0.05)。与含FBS的培养基相比,hBM-MSCs在含AS的培养基中培养时碱性磷酸酶活性显著更高(P < 0.05)。体内研究显示形成了类骨质样基质而非确定的骨组织。

结论

1)FBS适合hBM-MSCs的细胞增殖;2)含AS的培养基很可能有诱导hBM-MSCs分化的良好可能性;3)AS培养的hBM-MSCs包含一组在未进行骨诱导的情况下自发分化为类骨质样基质的细胞。

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