1. Accurate Determination of S-Phase Fraction in Proliferative Cells by Dual Fluorescence and Peroxidase Immunohistochemistry with 5-Bromo-2′-Deoxyuridine (BrdU) and Ki67 Antibodies
- Author
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Masao Doi, Takumi Ota, Hitoshi Okamura, Hiroyuki Kato, Shoichi Urabe, Naoki Mizuguchi, Motomi Tainaka, Jean-Michel Fustin, Rina Tanaka, Yoshiaki Yamaguchi, Shinshichi Hamada, and Yulin Chen
- Subjects
Male ,Pathology ,medicine.medical_specialty ,Histology ,Cellular differentiation ,Biology ,Article ,Antibodies ,S Phase ,chemistry.chemical_compound ,Mice ,Adrenal Glands ,medicine ,Gastric mucosa ,Animals ,Cell Proliferation ,Skin ,Wound Healing ,Staining and Labeling ,Cell growth ,Stomach ,Molecular biology ,Immunohistochemistry ,Epithelium ,Mice, Inbred C57BL ,medicine.anatomical_structure ,Ki-67 Antigen ,Antigen retrieval ,chemistry ,Bromodeoxyuridine ,Gastric Mucosa ,Indicators and Reagents ,Anatomy ,Wound healing - Abstract
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 G1-, S-, G2-, 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 G0 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.
- Published
- 2011