Juneja H S, Gardner F H, Minguell J J, Helmer R E
Exp Hematol. 1984 May;12(4):221-30.
Human bone marrow fibroblasts (BMF) were grown in vitro from normal (N) subjects and patients with aplastic anemia (AA). Growth studies in vitro revealed that both the N-BMF and AA-BMF had a logarithmic growth phase of eight days. Population doubling time for six of the 12 N-BMF and seven of the 12 AA-BMF was greater than 50 h. Five of the 12 AA-BMF studied in contrast to only two of the 12 N-BMF had a population doubling time of less than 50 h. The remaining four N-BMF had a population doubling time of greater than 100 h. During a similar duration in the logarithmic phase of growth, the AA-BMF underwent an average of 2.499 population doublings in comparison to 1.586 doublings by N-BMF (P = less than 0.01). The AA-BMF grew in multiple layers compared with the N-BMF, which usually grew as a monolayer. At the end of the logarithmic phase of growth, the AA-BMF also had a significantly higher number of cells per dish than the N-BMF (P = 0.02 on analysis of variance and covariance). These data suggest that a subgroup of AA-BMF grows faster than N-BMF and that the AA-BMF lack cell-to-cell inhibition. Testosterone, 3 alpha-etiocholanolone, and dexamethasone at 1 X 10(-8)M concentration, a physiological concentration, stimulated the growth of N-BMF as evidenced by increase in cell numbers and radioactive thymidine (3H-TdR) uptake. While dexamethasone had a stimulating effect on growth of N-BMF, it suppressed the growth of AA-BMF. Specific binding of radioactive dexamethasone (3H-dexa) was determined both for the N-BMF and AA-BMF. Specific binding sites for dexamethasone (Bmax) present on the N-BMF ranged from 460 to 770 fmol/mg protein). Bmax for AA-BMF was low (27-215 fmol/mg protein). In addition, the dissociation constant (Kd) was ten times lower for AA-BMF (1.0 X 10(-7) M) than for N-BMF (1.1 X 10(-8) M). The observations on the growth studies, the paradoxical response to dexamethasone, and the difference in the number of binding sites for dexamethasone indicate that the marrow fibroblasts from patients with aplastic anemia are abnormal.
从正常（N）受试者和再生障碍性贫血（AA）患者体内体外培养人骨髓成纤维细胞（BMF）。体外生长研究显示，N-BMF和AA-BMF均有8天的对数生长期。12个N-BMF中的6个以及12个AA-BMF中的7个群体倍增时间大于50小时。相比之下，12个研究的AA-BMF中有5个群体倍增时间小于50小时，而12个N-BMF中只有2个如此。其余4个N-BMF群体倍增时间大于100小时。在生长对数期的相似时间段内，AA-BMF平均经历2.499次群体倍增，而N-BMF为1.586次（P＜0.01）。与通常单层生长的N-BMF相比，AA-BMF呈多层生长。在生长对数期末，每培养皿中的AA-BMF细胞数量也显著多于N-BMF（方差分析和协方差分析P = 0.02）。这些数据表明，AA-BMF的一个亚组生长速度比N-BMF快，且AA-BMF缺乏细胞间抑制。1×10⁻⁸M浓度（生理浓度）的睾酮、3α-乙基胆甾烷醇酮和地塞米松刺激N-BMF生长，表现为细胞数量增加和放射性胸苷（³H-TdR）摄取增加。虽然地塞米松对N-BMF生长有刺激作用，但它抑制AA-BMF的生长。测定了N-BMF和AA-BMF对放射性地塞米松（³H-地塞米松）的特异性结合。N-BMF上存在的地塞米松特异性结合位点（Bmax）范围为460至770 fmol/mg蛋白质）。AA-BMF的Bmax较低（27 - 215 fmol/mg蛋白质）。此外，AA-BMF的解离常数（Kd）（1.0×10⁻⁷M）比N-BMF（1.1×10⁻⁸M）低10倍。关于生长研究的观察结果、对地塞米松的矛盾反应以及地塞米松结合位点数量的差异表明，再生障碍性贫血患者的骨髓成纤维细胞是异常的。
