Crisp A J, McGuire-Goldring M B, Goldring S R
Br J Exp Pathol. 1984 Dec;65(6):645-54.
Specimens of human trabecular bone from II patients were processed for tissue culture. In 10 out of 11 samples both cellular and matrix outgrowths were noted at the surfaces of explanted fragments after the first week in culture. During the second week adherent cells extended beyond the margins of the bone fragments and appeared to replicate. Plates achieved confluence in 30-36 days and cells were subcultured. In passaged cells doubling times were 5-7 days. Six cell cultures were examined for the presence of alkaline and acid phosphatase activity employing histochemical techniques. All six cultures contained cells which stained positively for alkaline phosphatase (10-80%). A small number of cells in one culture demonstrated tartrate-resistant acid phosphatase activity. Responses to hormones known to regulate the biological activities of skeletal tissues were also tested. Intracellular cyclic AMP was significantly increased by parathyroid hormone in three cultures, by salmon calcitonin in three cultures and by prostaglandin E2 in all 10 cultures. All three hormones increased the cyclic AMP content of cells cultured from human periosteum. It is concluded that cells cultured by this method demonstrate biochemical and morphological characteristics consistent with a skeletal tissue origin. Furthermore, such an approach may permit isolation and further characterization of individual subpopulations of bone cells of human origin.
对11例患者的人小梁骨标本进行组织培养处理。在11个样本中的10个样本中,培养第一周后,在外植碎片表面观察到细胞和基质生长。在第二周,贴壁细胞延伸到骨碎片边缘之外并似乎进行了复制。平板在30 - 36天达到汇合,细胞进行传代培养。传代细胞的倍增时间为5 - 7天。采用组织化学技术检查了6个细胞培养物中碱性磷酸酶和酸性磷酸酶活性的存在情况。所有6个培养物中均含有对碱性磷酸酶呈阳性染色的细胞(10% - 80%)。一种培养物中的少数细胞表现出抗酒石酸酸性磷酸酶活性。还测试了对已知调节骨骼组织生物活性的激素的反应。甲状旁腺激素使3个培养物中的细胞内环磷酸腺苷显著增加,鲑鱼降钙素使3个培养物中的细胞内环磷酸腺苷显著增加,前列腺素E2使所有10个培养物中的细胞内环磷酸腺苷显著增加。所有这三种激素均增加了从人骨膜培养的细胞的环磷酸腺苷含量。结论是,用这种方法培养的细胞表现出与骨骼组织起源一致的生化和形态学特征。此外,这种方法可能允许分离和进一步鉴定人源骨细胞的各个亚群。