Chen J L, Hunt P, McElvain M, Black T, Kaufman S, Choi E S
Amgen Inc., Thousand Oaks, California 91320, USA.
Stem Cells. 1997;15(5):368-77. doi: 10.1002/stem.150368.
It is known that osteoblast precursor cells are found in the low-density mononuclear (LDMN) fraction of human bone marrow (BM) aspirates. The purpose of this study was to investigate whether CD34, a hematopoietic progenitor cell marker, is present on osteoblast progenitor cells. LDMN, CD34+, and CD34- cells were cultured under conditions that promote growth and differentiation of mineral-secreting osteoblasts in a limiting dilution manner. With LDMN cells, osteoblast progenitor cells were found at an average frequency of 1/36,000 cells. With CD34- cells, osteoblast progenitor frequency remained at an average of 1/33,000, similar to LDMN cells. With CD34+ selected cells, osteoblast progenitor frequency increased to an average of 1/5,000. This osteoblast progenitor frequency is maintained in sorted CD34+/CD38+ cells. The osteoblasts generated from CD34+ cells were morphologically normal, and expression of skeletal-specific alkaline phosphatase and osteonectin increased upon differentiation induced by dexamethasone (DEX) treatment. Ultrastructurally, these CD34+ cell-derived osteoblasts displayed osteoblast-specific features. Functionally, these CD34+ cell-derived osteoblasts differentiated with DEX treatment, increased the level of cyclic adenosine monophosphate in response to parathyroid hormone stimulation, increased the level of alkaline phosphatase activity, and increased mineral secretion. These results demonstrate that osteoblast progenitor cells are enriched in the CD34+ cell population from BM and that these progenitor cells can differentiate into functional osteoblasts in culture.
已知在人骨髓抽吸物的低密度单核(LDMN)组分中可发现成骨细胞前体细胞。本研究的目的是调查造血祖细胞标志物CD34是否存在于成骨细胞前体细胞上。以有限稀释的方式在促进分泌矿物质的成骨细胞生长和分化的条件下培养LDMN细胞、CD34 +细胞和CD34 -细胞。对于LDMN细胞,成骨细胞前体细胞的平均频率为1/36,000个细胞。对于CD34 -细胞,成骨细胞祖细胞频率平均保持在1/33,000,与LDMN细胞相似。对于选择的CD34 +细胞,成骨细胞祖细胞频率增加到平均1/5,000。这种成骨细胞祖细胞频率在分选的CD34 + / CD38 +细胞中得以维持。由CD34 +细胞产生的成骨细胞形态正常,并且在用地塞米松(DEX)处理诱导分化后,骨骼特异性碱性磷酸酶和骨连接蛋白的表达增加。在超微结构上,这些源自CD34 +细胞的成骨细胞表现出成骨细胞特异性特征。在功能上,这些源自CD34 +细胞的成骨细胞经DEX处理后分化,响应甲状旁腺激素刺激而增加环磷酸腺苷水平,增加碱性磷酸酶活性水平,并增加矿物质分泌。这些结果表明,成骨细胞前体细胞在来自骨髓的CD34 +细胞群体中富集,并且这些祖细胞在培养中可分化为功能性成骨细胞。