Dennis J E, Merriam A, Awadallah A, Yoo J U, Johnstone B, Caplan A I
Skeletal Research Center, Department of Biology, Case Western Reserve University, Cleveland, Ohio 44106, USA.
J Bone Miner Res. 1999 May;14(5):700-9. doi: 10.1359/jbmr.1999.14.5.700.
Adult marrow contains mesenchymal progenitor cells (MPCs) that have multiple differentiation potentials. A conditionally immortalized MPC clone, BMC9, has been identified that exhibits four mesenchymal cell phenotypes: chondrocyte, adipocyte, stromal (support osteoclast formation), and osteoblast. The BMC9 clone, control brain fibroblasts and another marrow-derived clone, BMC10, were isolated from a transgenic mouse (H-2Kb-tsA58) containing a gene for conditional immortality. To test for chondrogenic potential, cells were cultured in defined medium containing 10 ng/ml transforming growth factor beta and 10-7 M dexamethasone in 15-ml polypropylene tubes ("aggregate cultures"). Adipogenic potential was quantitated by flow cytometry of Nile Red-stained cells cultured for 1 and 2 weeks in medium containing isobutyl methylxanthine, indomethacin, insulin, and dexamethasone. Support of osteoclast formation was measured by quantitating multinucleated tartrate-resistant acid phosphatase-positive cells in spleen cell cocultures of test clones (immortomouse clones and positive control ST2 cells) cultured in the presence of 10-7 M vitamin D3 and 150 mM ascorbate-2-phosphate. In vivo osteogenic potential was assayed by histologic examination of bone formation in subcutaneous implants, into athymic mouse hosts, of a composite of cells combined with porous calcium phosphate ceramics. The bone marrow-derived clone BMC9 has the potential to express each of the four mesenchymal characteristics tested, while brain fibroblasts, tested under identical conditions, did not exhibit any of these four mesenchymal characteristics. BMC10 cells exhibited osteogenic and chondrogenic phenotypes, but showed only minimal expression of adipocytic or osteoclast-supportive phenotypes. Clone BMC9 is, minimally, a quadripotential MPC isolated from the marrow of an adult mouse that can differentiate into cartilage and adipose, support osteoclast formation, and form bone. The BMC9 clone is an example of an adult-derived multipotential progenitor cell that is situated early in the mesenchymal lineage.
成人骨髓含有具有多种分化潜能的间充质祖细胞（MPC）。已鉴定出一种条件永生化的MPC克隆BMC9，它表现出四种间充质细胞表型：软骨细胞、脂肪细胞、基质细胞（支持破骨细胞形成）和成骨细胞。BMC9克隆、对照脑成纤维细胞和另一个骨髓来源的克隆BMC10是从一只含有条件永生化基因的转基因小鼠（H-2Kb-tsA58）中分离出来的。为了测试软骨形成潜能，将细胞在含有10 ng/ml转化生长因子β和10-7 M地塞米松的限定培养基中，于15 ml聚丙烯管中培养（“聚集培养”）。通过对在含有异丁基甲基黄嘌呤、吲哚美辛、胰岛素和地塞米松的培养基中培养1周和2周的尼罗红染色细胞进行流式细胞术来定量脂肪生成潜能。通过在10-7 M维生素D3和150 mM抗坏血酸-2-磷酸存在下培养的测试克隆（永生小鼠克隆和阳性对照ST2细胞）的脾细胞共培养物中定量多核抗酒石酸酸性磷酸酶阳性细胞来测量对破骨细胞形成的支持。通过对皮下植入无胸腺小鼠宿主的细胞与多孔磷酸钙陶瓷复合物的骨形成进行组织学检查来测定体内成骨潜能。骨髓来源的克隆BMC9具有表达所测试的四种间充质特征中每一种的潜能，而在相同条件下测试的脑成纤维细胞未表现出这四种间充质特征中的任何一种。BMC10细胞表现出成骨和软骨形成表型，但仅表现出极少量的脂肪细胞或支持破骨细胞表型的表达。克隆BMC9至少是从成年小鼠骨髓中分离出的一种四能MPC，它可以分化为软骨和脂肪、支持破骨细胞形成并形成骨。BMC9克隆是位于间充质谱系早期的成年来源多能祖细胞的一个例子。
