Differential expression of alkaline phosphatase in clones of human osteoblast‐like cells
Differential expression of alkaline phosphatase in clones of human osteoblast‐like cells
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人成骨细胞样细胞克隆中碱性磷酸酶的差异表达
DOI：
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发表时间：
1993
影响因子：
6.2
通讯作者：
A. Federici
中科院分区：
文献类型：
--
作者：
P. Manduca;C. Sanguineti;M. Pistone;E. Boccignone;F. Sanguineti;F. Santolini;A. Federici
我们建立了从成人骨片中生长出来的细胞培养，并在培养中维持了12代。培养显示成骨细胞表型，并伴有I型胶原、骨连接素、碱性磷酸酶和骨钙素的合成。我们报告了从三个不同的个体原代培养中获得的21个克隆的特性。我们研究了骨连接素、碱性磷酸酶、胶原蛋白和骨钙素在克隆中的表达。代谢标记显示，在所研究的所有克隆中都产生了I型胶原蛋白和骨连接素。在三分之二的无性系和大量培养中，在传代2时未检测到碱性磷酸酶，但在培养的后期传代中检测到碱性磷酸酶的数量不断增加。通过传代8，克隆获得了不同但可检测的表达水平。碱性磷酸酶在阳性克隆中的表达水平不同，可能是因为它们来源于成骨细胞成熟的不同阶段，也可能是由于微环境的微小变化。碱性磷酸酶阳性克隆检测骨钙素，仅在传代10时可测量表达。经12代培养，获得的无性系中有1 / 3碱性磷酸酶呈阴性。获得不能产生碱性磷酸酶的克隆可能是由于在离体培养条件下失去了分化潜能。无论碱性磷酸酶的表达潜力如何，所有克隆的生长速度和潜力在培养12代时都是相似的。
We established cultures of cells growing out from adult bone chips and maintained them through 12 passages in culture. The cultures showed osteoblastic phenotype accompanied by synthesis of collagen type I, osteonectin, alkaline phosphatase, and osteocalcin. We report the chracterization of 21 clones obtained from three different individual primary cultures. We studied the expression of osteonectin, alkaline phosphatase, collagen, and osteocalcin in the clones. Metabolic labeling showed production of type I collagen and of osteonectin in all clones studied. In two‐thirds of the clones and in mass cultures alkaline phosphatase was not detected at passage 2, but it was detected in increasing amounts at later passages in culture. The clones attained different but detectable levels of expression of this marker by passage 8. The different levels in the expression of alkaline phosphatase in positive clones may be because they were derived from cells at different stages of osteoblastic maturation or due to small changes in microenvironment. The alkaline phosphatase‐positive clones were tested for osteocalcin, and they showed measurable expression only at passage 10. A third of the clones obtained were negative for alkaline phosphatase during 12 passages in culture. The obtainment of clones unable to produce alkaline phosphatase may be due to loss of differentiating potential under the in vitro culture conditions. The growth rate and potential of all clones studied were similar through 12 passages in culture, regardless of their potential for expression of alkaline phosphatase.
影响因子：
2.7
作者：
GERSTENFELD, LC;CHIPMAN, SD;LIAN, JB
通讯作者：
LIAN, JB