DOUBLE LABELING WITH IODODEOXYURIDINE AND BROMODEOXYURIDINE FOR CELL-KINETICS STUDIES
DOUBLE LABELING WITH IODODEOXYURIDINE AND BROMODEOXYURIDINE FOR CELL-KINETICS STUDIES
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DOI：
10.1177/37.7.2659659
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发表时间：
1989-07-01
影响因子：
3.2
通讯作者：
GRAY, JW
中科院分区：
文献类型：
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作者：
SHIBUI, S;HOSHINO, T;GRAY, JW
通过细胞周期的进展率测定在5个人胶质瘤细胞系通过一种新的顺序免疫组化染色技术。首先用碘脱氧尿苷（IdUrd）标记细胞1-3小时，然后用溴脱氧尿苷（BrdUrd）标记30分钟。用Br-3（一种仅识别BrdUrd的单克隆抗体）和IU-4（一种识别IdUrd和BrdUrd的抗体）鉴定标记细胞。每个载玻片依次染色，首先用免疫过氧化物酶法染色Br-3，然后用碱性磷酸酶-抗碱性磷酸酶法染色IU-4。仅对IU-4呈阳性的细胞代表在与IdUrd孵育期间进入G2期的S期细胞的分数。通过该方法测量的进展速率在每个细胞系中是恒定的，并且导致比通过在不同载玻片中单独对IdUrd和BrdUrd染色的标本的测量获得的标准误差更小的标准误差。 在5种细胞系中，从该组分计算的S期持续时间范围为8-13小时;估计的潜在倍增时间为25-32小时，与实际倍增时间非常相似。
The rate of progression through the cell cycle was determined in five human glioma cell lines by a new sequential immunohistochemical staining technique. The cells were labeled first with iododeoxyuridine (IdUrd) for 1-3 hr and then with bromodeoxyuridine (BrdUrd) for 30 min. Labeled cells were identified with Br-3, a monoclonal antibody that recognizes only BrdUrd, and with IU-4, an antibody that recognizes both IdUrd and BrdUrd. Each slide was stained sequentially, first with the immunoperoxidase method for Br-3 and then with the alkaline phosphatase-anti-alkaline phosphatase method for IU-4. Cells that were positive only for IU-4 represented the fraction of S-phase cells that passed into the G2 phase during the period of incubation with IdUrd. The rates of progression measured by this method were constant in each cell line and resulted in smaller standard errors than were obtained by measurements from specimens stained singly for IdUrd and BrdUrd in different slides. The duration of the S-phase calculated from this fraction in the five cell lines ranged from 8-13 hr; the estimated potential duoubling times were 25-32 hr and were very similar to the actual doubling times.