EVIDENCE FOR HETEROGENEITY OF THE OSTEOBLASTIC PHENOTYPE DETERMINED WITH CLONAL RAT BONE-CELLS ESTABLISHED FROM TRANSFORMING GROWTH FACTOR-BETA-INDUCED CELL COLONIES GROWN ANCHORAGE INDEPENDENTLY IN SEMISOLID MEDIUM
EVIDENCE FOR HETEROGENEITY OF THE OSTEOBLASTIC PHENOTYPE DETERMINED WITH CLONAL RAT BONE-CELLS ESTABLISHED FROM TRANSFORMING GROWTH FACTOR-BETA-INDUCED CELL COLONIES GROWN ANCHORAGE INDEPENDENTLY IN SEMISOLID MEDIUM
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DOI：
10.1210/endo-125-4-2092
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发表时间：
1989-10-01
期刊：
影响因子：
4.8
通讯作者：
FLEISCH, H
中科院分区：
文献类型：
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作者：
GUENTHER, HL;HOFSTETTER, W;FLEISCH, H
文献中出现了许多报告，表明成骨细胞谱系细胞之间存在表型异质性。这种多样性可能是由于分化成骨细胞的某些阶段。为了获得这个问题的答案，我们报告的研究进行克隆骨细胞群从出生后1天大鼠颅骨表达明确定义的表型差异。为了实现这一目标，我们使用了软琼脂克隆技术，以前几乎只适用于肿瘤起源的细胞克隆。能够采用该方法的原因是基于骨细胞可以被转化生长因子-β诱导的事实。可逆地获得转化表型，这是由锚定依赖性骨细胞表达的事件，以在软琼脂糖中形成逐渐生长的集落。将从琼脂糖收获的单个集落扩增为克隆骨细胞群。通过检测成骨细胞标志物如碱性磷酸酶活性来表征48个细胞克隆。PTH和E2诱导的腺苷酸环化酶活性，骨钙素mRNA合成，以及胶原合成，成骨细胞类型的7个子集被确定。发现每个亚群表达不同的表型，由成骨细胞标志物的存在或缺失指示。以前发现不显示任何成骨细胞特性的一些克隆，当用胰岛素样生长因子-I/转化生长因子-β处理时，提示这些克隆可能来源于骨祖细胞库。虽然大多数克隆细胞群体的特征在于作为完全功能的成骨细胞，一些克隆只表达1，2，或3成骨细胞标志物，这表明它们可能代表单独的成骨途径的分化阶段。此外，其他亚克隆显示合成骨钙素的能力，并显示PTH和肾上腺素-E2的反应，但被发现是缺乏碱性磷酸酶活性。其他人表达所有成骨细胞标志物，除了PTH反应。后者的表型星座表明，这些细胞克隆可能代表成熟的成骨细胞样细胞，这可能是由于在分离时存在的环境条件，已成为根据组织的生理要求改变。
Numerous reports have appeared in the literature indicating phenotypic heterogeneity among cells of the osteoblastic lineage. This diversity may be due to either certain stages of differentiated osteoblasts. To obtain answers to this question, we report on studies undertaken to clone bone cell populations from 1 day postnatal rat calvaria which express well defined differences in phenotype. To achieve this goal, we have used the soft agarose cloning technique which previously has almost exclusively been applied to clone cells of neoplastic origin. The reason for being able to employ this method is based on the fact that bone cells can be induced by transforming growth factor-.beta. to reversibly acquire the transformed phenotype, an event expressed by anchorage-dependent bone cells to form progressively growing colonies in soft agarose. Individual colonies, harvested from agarose, were expanded to clonal bone cell populations. Characterizing 48 cell clones by detection of osteoblastic cell markers such as alkaline phosphatase activity. PTH- and prostaglandin-E2-induced adenylate cyclase activity, osteocalcin mRNA synthesis, as well as collagen synthesis, 7 subsets of osteoblastic cell types were identified. Each subset was found to express a distinct phenotype, indicated by the absence or presence of osteoblastic cell markers. Some clones, previously found not to exhibit any osteoblastic traits, developed PTH responsiveness when treated with insulin-like growth factor-I/transforming growth factor-.beta., suggesting that these clones may originate from the osteoprogenitor cell pool. While most clonal cell populations were characterized as fully functional osteoblastic cells, some clones expressed merely 1, 2, or 3 osteoblastic markers, which suggests that they may represent stages of differentiation alone the osteogenic pathway. In addition, other subclones displayed the capacity to synthesize osteocalcin and showed PTH and prostaglandin-E2 responsiveness, but were found to be devoid of alkaline phosphatase activity. Others expressed all osteoblastic cell markers except PTH responsiveness. The phenotype constellation of the latter suggest that these cell clones may represent mature osteoblast-like cells, which, perhaps due to environmental circumstances present at the time of isolation, have become altered in accordance with the physiological requirements of the tissue.