In Vitro Study of Osteogenesis from Isolated Light Density Bone Marrow Cells

小比重骨髓細胞體外骨生成的研究

史中(C Shih) ； 張燕清(Y C Chang) ； 石淦生(K S Shih) ； 彭志綱(T K Peng) ； 王天美(T M Wang)

《中華民國口腔顎面外科學會雜誌》 13卷1期 (2002/03) Pp. 1-9

https://doi.org/10.7104/CJOMS.200203.0001

摘要

造骨細胞源自存在骨髓中數前身細胞。由於一直無法自骨髓細胞培養過程之貼附細胞分離出造骨原始細胞。以往對骨髓細胞的探討，常常是使用已分化之造骨原細胞且其中含有所有的血源性細胞。本研究旨在利用小比重骨髓細胞發展出適合探討骨生成之細胞培養模式。利用自八週大BALB/C雄性小白鼠股骨和脛骨獲得之小比重骨髓細胞，並使用BGJb培養液培養。結果發現：小比重骨髓細胞培養４天後，可見未礦物化群體形成。6-9天，經von Kossa染色法則有礦物化骨群體形成，並且骨群體之超微結構與體內骨生成之超微結構相似。骨群體形成的數目和植入的細胞呈現正相關，表示造骨幹細胞能夠在體外貼附、增生、分化並形成骨群體。另外，隨著植入細胞數目的增加，鹼性磷酸活性呈現明顯地增加。因此，本研究所發展出的骨髓細胞培養模式適合於進行骨生成細胞增生、分化的和礦物化過程調控之相關研究。

關鍵字

骨群體 ； 骨髓細胞 ； 分化 ； 增生 ； 骨生成

並列摘要

Osteoblasts are derived from progenitors present in low frequency in bone marrow, Because of the lack of a practical procedure to isolate osteoblast progenitors from early culture of adherent cells from bone marrow, previous studies of marrow cells have been made in confluent cultures of bone marrow when the osteoblast progenitors are already differentiated. These studies utilized cultures containing mixed populations of cells including all the hemotopoiatic cells. The purpose of this study was to develop a suitable in vitro culture system of osteogenesis using light density bone marrow cells. Light density bone marrow cells were obtained from femur and tibiae of 8-week -old BALB/c male mice. After culturing light density bone marrow cells for 4 days, unmineralized colonies began to form in culture dish. When culturing for anther 6to 9days, the osteogenic colonies were identified by von Kossa staining and had the between the number of osteogenic colonies formed in dishes and the number of cells plated, inicating a primitive stem cell capable colony. In addition, significant increase in alkaline phosphatase activity was positively correlated to the cell plated. In conclusion, a suitable in vitro bone marrow cell culture system can be developed for the study of factors regulating proliferation and differentiation of osteogenic lineage cell, and formation of mineralized bone.

並列關鍵字

Bone colony ； Bone marrow cells ； Differentiation ； Proliferation ； Osteogenesis