Casasco A, Casasco M, Cornaglia A I, Mazzini G, De Renzis R, Tateo S
Institute of Histology and Embryology, University of Pavia, Italy.
Connect Tissue Res. 1995;32(1-4):63-70. doi: 10.3109/03008209509013707.
The development of antibodies to cell cycle-related antigens provides the basis for immunochemical studies on cell kinetics. Bromo-deoxyuridine (BrdU) incorporated by S-phase traversing cells is an exogenous marker of replicating cells, whereas proliferating cell nuclear antigen (PCNA) is an endogenous marker of replicating cells. We have applied monoclonal antibodies to BrdU and PCNA to study cell kinetics in tooth germ by immunohistochemistry and flow cytometry. BrdU-antibody reacted only with S phase-traversing cells in pulse-labelling experiments, whereas PCNA-antibody reacted with G1, S and G2-M phases traversing cells. Although the number of PCNA-positive cells largely exceeded the number of BrdU-labelled cells, the pattern distribution of immunoreactive cells was similar using BrdU- and PCNA-antibodies as revealed by immunohistochemistry. The use of PCNA-antibody allowed the detection of proliferating cells also in human tooth germ. It is suggested that combined identification of BrdU and PCNA on one side and growth factors, oncoproteins or differentiation markers on the other side may constitute a useful approach to understand the mechanisms of cell differentiation in tooth germ.
针对细胞周期相关抗原的抗体的发展为细胞动力学的免疫化学研究提供了基础。处于S期的细胞掺入的溴脱氧尿苷（BrdU）是复制细胞的外源性标记物，而增殖细胞核抗原（PCNA）是复制细胞的内源性标记物。我们已应用针对BrdU和PCNA的单克隆抗体，通过免疫组织化学和流式细胞术研究牙胚中的细胞动力学。在脉冲标记实验中，BrdU抗体仅与处于S期的细胞发生反应，而PCNA抗体则与处于G1、S和G2 - M期的细胞发生反应。尽管PCNA阳性细胞的数量大大超过BrdU标记细胞的数量，但免疫组织化学显示，使用BrdU和PCNA抗体时，免疫反应性细胞的模式分布相似。使用PCNA抗体也能够检测人牙胚中的增殖细胞。有人提出，一方面联合鉴定BrdU和PCNA，另一方面联合鉴定生长因子、癌蛋白或分化标记物，可能构成一种了解牙胚细胞分化机制的有用方法。
