Bromodeoxyuridine을 사용한 세포동시화와 후기 DNA복제

초록

Recent advances in cytogentic technology have improved the resolving power of cytogenetic diagnosis. High resoultion banding by the study of the less contracted chromosomes with synchronization techniques became useful in refined studies of chromosome ultrastructure and in the detection of minute chromosomal changes. A simple method using bromodeoxyuridine (BrdU), for both cell synchronization and incorporation into replicating DNA is described. PHA-stimulated peripheral lymphocytes from chromosomally normal subjects were cultures, synchronized and incorporated by BrdU (200㎍/㎖)for 15-17 hours, and pulsed with thymidine (0.3㎍/㎖)for 6 hours. The results are summarized as follows: 1. The late replicating elements were G-bands, C-bands regions. 2. The mitotic index was 9.2％ 3. The cells were at various stages of chromosome contraction, but prometaphase and early metaphase were predominant. This study shows that this methodology is a potentially valuable clinical cytogenetic technique, because it: 1)produce sharp band definition and increased resolution in metaphase and prometaphase cells of the same preparation, 2) is that the mitotic index was high.