Park S R, Oreffo R O, Triffitt J T
MRC Bone Research Laboratory, University of Oxford, Nuffield Orthopaedic Centre, UK.
Bone. 1999 Jun;24(6):549-54. doi: 10.1016/s8756-3282(99)00084-8.
Information on the interconversion potential of adipocytes and other end cells characteristic of the stromal fibroblastic cell lineages, key in the understanding of bone turnover in metabolic diseases such as osteoporosis, is limited. The object of the present study was: i) to isolate relatively pure populations of adipocytes from human bone marrow; ii) to clone single adipocytes from these populations; and iii) to examine in vitro the interconversion potential of the progeny of these single-cloned adipocytes between the osteogenic and adipogenic phenotypes. Adipogenic colonies were isolated from the low-density floating fraction of normal bone marrow cells cultured in adipogenic media for 4 days. Single adipocytes were isolated and cloned by limiting dilution. Cloned adipocytes were found to dedifferentiate into fibroblast-like cells, and subsequently to differentiate into two morphologically distinct cell types: osteoblasts and adipocytes in appropriate culture systems. The adipocytic phenotype was confirmed by morphology, oil red O staining, and immunocytochemistry using antiserum to aP2. The osteogenic phenotype was confirmed by alkaline phosphatase, osteocalcin immunostaining using specific osteocalcin antiserum, and formation of mineralized cell aggregates. These findings demonstrate the extent of plasticity between the differentiation of adipocytic and osteogenic cells in human bone marrow stromal cell cultures. We have shown the ability of isolated clonal adipogenic cells to redifferentiate into cells of the osteogenic and adipogenic lineage and the interconversion potential of human marrow stromal cells in vitro. These results provide further evidence that the osteogenic and adipogenic cells share a common multipotential precursor.
关于脂肪细胞与其他属于基质成纤维细胞谱系的终末细胞之间相互转化潜能的信息有限，而这对于理解骨质疏松等代谢性疾病中的骨转换至关重要。本研究的目的是：i）从人骨髓中分离出相对纯净的脂肪细胞群体；ii）从这些群体中克隆单个脂肪细胞；iii）在体外检测这些单克隆脂肪细胞后代在成骨和成脂表型之间的相互转化潜能。在成脂培养基中培养4天的正常骨髓细胞的低密度漂浮部分分离出成脂集落。通过有限稀释法分离并克隆单个脂肪细胞。发现克隆的脂肪细胞去分化为成纤维样细胞，随后在适当的培养系统中分化为两种形态不同的细胞类型：成骨细胞和脂肪细胞。通过形态学、油红O染色以及使用抗aP2抗血清的免疫细胞化学方法证实了脂肪细胞表型。通过碱性磷酸酶、使用特异性骨钙素抗血清的骨钙素免疫染色以及矿化细胞聚集体的形成证实了成骨细胞表型。这些发现证明了人骨髓基质细胞培养物中脂肪生成和成骨细胞分化之间的可塑性程度。我们已经展示了分离的克隆成脂细胞重新分化为成骨和成脂谱系细胞的能力以及人骨髓基质细胞在体外的相互转化潜能。这些结果进一步证明成骨细胞和成脂细胞共享一个共同的多能前体。
