Accurate Determination of S-Phase Fraction in Proliferative Cells by Dual Fluorescence and Peroxidase Immunohistochemistry with 5-Bromo-2 '-Deoxyuridine (BrdU) and Ki67 Antibodies

作者:Tanaka Rina; Tainaka Motomi; Ota Takumi; Mizuguchi Naoki; Kato Hiroyuki; Urabe Shoichi; Chen Yulin; Fustin Jean Michel; Yamaguchi Yoshiaki; Doi Masao; Hamada Shinshichi; Okamura Hitoshi*
来源:Journal of Histochemistry and Cytochemistry, 2011, 59(8): 791-798.
DOI:10.1369/0022155411411090

摘要

To ensure the maintenance of tissues in mammals, cell loss must be balanced with cell production, the proliferative activity being different from tissue to tissue. In this article, the authors propose a new method for the quantification of the proliferative activity, defined as the S-phase fraction of actively cycling cells, by dual labeling with fluorescence and peroxidase immunohistochemistry using BrdU (marker of S-phase) and Ki67 antibodies (marker of G(1)-, S-, G(2)-, and M-phases) after a one-step antigen retrieval. In the generative cell zones of fundic and pyloric glandular stomachs, where the majority of cells were cycling, the authors measured a proliferative activity of 31%. In the epithelium of the forestomach and the skin, where cycling cells are intermingled with G(0) and differentiated cells, proliferative activities were 21% and 13%, respectively. In the adrenal cortex, in which cycling cells were sparsely distributed, the proliferative activity reached 32%. During the regenerative process in the skin after a lesion, the proliferative activity increased in proximity to the wound. The present one-step dual-labeling method has revealed that the proliferative activity is different between tissues and depends on the physiological or pathological state. (J Histochem Cytochem 59:791-798, 2011)

  • 出版日期2011-8