Takeda Akira, Yamazaki Yasuharu, Baba Kyoko, Ishiguro Masashi, Aoyagi Kazuya, Ikemoto Shigehiro, Uchinuma Eiju
Department of Plastic and Aesthetic Surgery, School of Medicine, Kitasato University, Kanagawa, Japan.
J Oral Maxillofac Surg. 2012 Aug;70(8):e469-76. doi: 10.1016/j.joms.2012.03.021.
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.
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.
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.
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。
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。
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。
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。