Nakahara H, Dennis J E, Bruder S P, Haynesworth S E, Lennon D P, Caplan A I
Department of Biology, Case Western Reserve University, Cleveland, Ohio 44106.
Exp Cell Res. 1991 Aug;195(2):492-503. doi: 10.1016/0014-4827(91)90401-f.
Periosteal cells were enzymatically liberated from the tibiae of young chicks, introduced into cell culture, and allowed to reach confluence. The morphology of the cells gave the impression of a relatively homogeneous population of fibroblast-like cells. These cultured cells did not overtly express osteogenic or chondrogenic properties as judged by their morphology and the lack of reactivity with probes to phenotype-specific antigens of osteoblasts or chondrocytes. The cells were then replated at relatively high density and chronologically evaluated for the differentiation of bone and cartilage. These replated cells formed a multi-layer of fibroblast-like cells, the top portion of which eventually differentiated into bone tissue as evidenced by the presence of mineralization and immunocytochemical reactivity to bone Gla protein- and osteocyte-specific probes. Cells below this distinctive top layer differentiated into chondrocytes, which eventually further developed into hypertrophic chondrocytes as evidenced by their morphology and the presence of immunoreactive type X collagen in the matrix. Mineralization was also observed in the territorial matrix of these hypertrophic chondrocytes, when the culture was augmented with beta-glycerophosphate. Periosteal-derived cells replated at a lower density as controls did not show signs of osteochondrogenic differentiation. These observation suggest that periosteal-derived cells of young chicks contain mesenchymal cells which possess the potential to undergo terminal differentiation into osteogenic or chondrogenic phenotypes depending on local environmental or positional cues.
将幼年雏鸡胫骨中的骨膜细胞通过酶解分离出来，接种到细胞培养物中，使其达到汇合状态。这些细胞的形态给人一种相对均匀的成纤维细胞样细胞群体的印象。从形态以及与成骨细胞或软骨细胞表型特异性抗原探针缺乏反应性判断，这些培养的细胞未明显表达成骨或成软骨特性。然后将这些细胞以相对较高的密度重新接种，并按时间顺序评估其向骨和软骨的分化情况。这些重新接种的细胞形成了多层成纤维细胞样细胞，其顶部最终分化为骨组织，这可通过矿化的存在以及对骨钙素和骨细胞特异性探针的免疫细胞化学反应来证明。在这个独特顶层下方的细胞分化为软骨细胞，最终进一步发育为肥大软骨细胞，这可从其形态以及基质中存在免疫反应性X型胶原得到证明。当培养物中添加β-甘油磷酸时，在这些肥大软骨细胞的区域基质中也观察到了矿化现象。作为对照，以较低密度重新接种的骨膜来源细胞未显示出骨软骨分化的迹象。这些观察结果表明，幼年雏鸡的骨膜来源细胞含有间充质细胞，这些细胞具有根据局部环境或位置线索进行终末分化为成骨或成软骨表型的潜力。
