Nakahara H, Bruder S P, Haynesworth S E, Holecek J J, Baber M A, Goldberg V M, Caplan A I
Department of Biology, Case Western Reserve University, Cleveland, Ohio.
Bone. 1990;11(3):181-8. doi: 10.1016/8756-3282(90)90212-h.
Periosteal cells were enzymatically isolated from the tibiae of young chicks, introduced into cell culture, allowed to reach confluence, and subcultured. The freshly isolated or subcultured cells were loaded into diffusion chambers and implanted into the peritoneal cavity of athymic mice to test their osteo-chondrogenic potential in a contained in vivo location. Freshly isolated periosteal cells formed both bone and cartilage tissue in such test chambers, but with a relatively low incidence. In contrast, cultured periosteal cells consistently gave rise to bone and cartilage even after 10 population doublings. With further passages of cells, the osteo-chondrogenic potential diminished substantially, until complete loss of expressivity at 16 population doublings or longer. Cultured muscle fibroblasts, when loaded into diffusion chambers under identical conditions to those of cultured periosteal cells, formed neither bone nor cartilage. These observations suggest that periosteal cells of young chicks contain subsets of progenitor cells or mesenchymal stem cells which possess the potential to differentiate into osteoblasts or chondrocytes, and this potential is retained after enzymatic isolation and for several population doublings in culture.
从幼雏胫骨中酶解分离出骨膜细胞，接种到细胞培养物中，待其达到汇合状态后进行传代培养。将新鲜分离或传代培养的细胞装入扩散小室，植入无胸腺小鼠的腹腔，以在体内限定部位测试其成骨软骨生成潜能。新鲜分离的骨膜细胞在这种测试小室中形成了骨和软骨组织，但发生率相对较低。相比之下，培养的骨膜细胞即使在经过10次群体倍增后仍能持续产生骨和软骨。随着细胞进一步传代，成骨软骨生成潜能大幅降低，直至在16次群体倍增或更长时间后完全丧失表达能力。在与培养的骨膜细胞相同的条件下将培养的肌肉成纤维细胞装入扩散小室，其既不形成骨也不形成软骨。这些观察结果表明，幼雏的骨膜细胞含有祖细胞或间充质干细胞亚群，它们具有分化为成骨细胞或软骨细胞的潜能，并且这种潜能在酶解分离后以及在培养中经过几次群体倍增后仍能保留。
