Hozier J C, Lindquist L L
Hum Genet. 1980 Feb;53(2):205-9. doi: 10.1007/BF00273497.
We have developed a new protocol for the preparation of banded chromosomes from human bone marrow. This protocol incorporates new procedures with improvements in conventional ones to rapidly produce high quality banded karyotypes from bone marrow aspirates. Tissue culture is completely eliminated and replaced with a truly direct method of chromosome preparation in which a small amount of marrow is treated with a solution containing trypsin, hypotonic salts and colcemid (THC). The THC protocol, when compared with standard short term culture methods for marrow chromosome preparation, produces more extended and more readily banded chromosomes. Rapid banding is further facilitated by replacement of standard G-banding technique with Wright's staining. The technical developments allow karyotypic analysis within 2-4 h after receipt of the specimen. The high quality and rapidity of the THC protocol have important implications for the clinical usefulness of cytogenetic analysis of bone marrow in studying congenital defects as well as leukemias and lymphomas.
我们已经开发出一种用于制备人类骨髓带型染色体的新方案。该方案结合了新程序并改进了传统程序,以便从骨髓抽吸物中快速产生高质量的带型核型。完全省去了组织培养,代之以一种真正直接的染色体制备方法,即使用含有胰蛋白酶、低渗盐和秋水仙酰胺(THC)的溶液处理少量骨髓。与用于骨髓染色体制备的标准短期培养方法相比,THC方案能产生更长且更容易显带的染色体。用瑞氏染色取代标准G显带技术进一步促进了快速显带。这些技术进展使在收到标本后2至4小时内即可进行核型分析。THC方案的高质量和快速性对于骨髓细胞遗传学分析在研究先天性缺陷以及白血病和淋巴瘤方面的临床应用具有重要意义。