Background: The size of the germinative growth fraction (i.e. the number of actively proliferating germinative cells) of normal human epidermis is still a subject of debate. Ki-67 antigen and PCNA, an auxiliary protein of d-polymerase, are considered as markers of the growth fraction when used under optimal conditions. Method: In the present work, we have compared Ki-67 expression (detected with MIB1 antibody) with PCNA expression (detected with PC10 antibody) in biopsies of normal human epidermis fixed in neutral formalin and using antigen retrieval by microwave processing. To obtain additional information, such as the percentage of cells in S phase, biopsies were also incubated in 3H-thymidine before immunostaining. Results: Before microwave treatment, 8% of the basal cells were positive for MIB1 antibody and 7.8% were positive for PC10 antibody. The 3H-thymidine labelling index was 2.8%. The proportion of MIB1-positive cells rose to 19% after antigen retrieval by microwave processing. In the same way, the 3H labelling index rose to 9%. In contrast, PC10 became positive in all epidermal nuclei. Conclusion: These results suggest that the growth fraction of the germinative cell population of normal human epidermis is not larger than 20% and is composed of cells with a short cell cycle time.

1.
Heenen M, Galand P: The growth fraction of normal human epidermis. Dermatology 1997; 194:313–317.
2.
Gerdes J, Lemke H, Wacker HH, Schwab J, Stein H: Cell cycle analysis of a cell proliferation associated human nuclear antigen defined by the monoclonal antibody Ki 67. J Immunol 1984;133:1710–1715.
3.
Brown DC, Gatter KC: Monoclonal antibody Ki 67: Its use in histopathology. Histopathology 1990;17:489–503.
4.
Gerdes J, Beker MHG, Key G, Cattoreti G: Immunological detection of tumor growth fraction (Ki 67 antigen) in formalin fixed and routinely processed tissue. J Pathol 1992;168: 85–87.
5.
Bravo R, Frank R, Blundel PA, MacDonald-Bravo H: Cyclin/PCNA is the auxiliary protein of DNA polymerase d. Nature 1987;326: 515–517.
6.
Van Dierendonck JH, Wijsman JH, Keijzer R, Vandevelde CJH, Cornelisse CJ: Cell cycle related staining pattern of antiproliferating cell nuclear antigen monoclonal antibody. Am J Pathol 1991;138:1165–1172.
7.
Wada T, Shimabukuro T, Matsuyama H, Naito K, Skog S, Tribukat B: Optimal conditions of fixation for immunohistochemical staining of proliferating cell nuclear antigen in tumor cells and its cell cycle immunohistochemical expression. Cell Prolif 1994;27:541–551.
8.
Galand P, Degraef CH: Cyclin/PCNA immunostaining as an alternative to tritiated thymidine pulse labelling for marking S phase cells in paraffin sections from animal and human tissues. Cell Tissue Kinet 1989;22:383–392.
9.
Morita T, Usuda N, Hanai T, Nagata T: Changes of colon epithelium proliferation due to individual aging with cyclin proliferating cell nuclear antigen (PCNA/cyclin) immunostaining compared to 3H-thymidine radioautoradiography. Histochemistry 1994;101:13–20.
10.
Celis JE, Celis A: Cell cycle dependent variations in the distribution of the nuclear protein cyclin/proliferating cell nuclear antigen in cultured cells: Subdivisions of S phase. Proc Natl Acad Sci USA 1985;82:3262–3266.
11.
Berlingin E, Heenen M, Galand P: Measurement of S phase duration in human epidermis using cyclin immunostaining and 3H-thymidine pulse labelling. Arch Dermatol Res 1992; 284:238–241.
12.
Kubbutat MHG, Cattoreti G, Gerdes J, Key G: Comparison of monoclonal antibodies PC10 and MIB1 on microwave processed paraffin sections. Cell Prolif 1994;27:553–559.
13.
Miyagawa S, Okada N, Takasaki Y, Takashi Y, Kitano Y, Yoshikawa K, Sakamoto K, Steinberg ML: Expression of proliferating cell nuclear antigen/cyclin in human keratinocytes. J Invest Dermatol 1989;93:678–681.
14.
Penneys NS, Bogaert M, Serfling U, Sisto M: PCNA expression in cutaneous keratinous neoplasm and verruca vulgaris. Am J Pathol 1992; 141:139–142.
15.
Furukawa F, Imamura S, Fujita M, Kinoshita K, Yoshitake K, Brown WR, Norris DA: Immunohistochemical location of proliferating cell nuclear antigen/cyclin in human skin. Arch Dermatol Res 1992;284:86–91.
16.
Van Erp PEJ, De Mare S, Rijzewijk JJ, Van de Kerckhof PCM, Bauer FW: A sequential double immunoenzymic staining procedure to obtain cell kinetic information in normal and hyperproliferative epidermis. Histochem J 1989; 21:343–347.
17.
Gerritsen JP, Van Erp PEJ, Van Vlijmen-Willems IMJJ, Lenders LTM, Van de Kerckhof PCM: Repeated tape stripping of normal skin: A histological assessment and comparison with events seen in psoriasis. Arch Dermatol Res 1994;286:455–461.
18.
Van Erp PEJ, Groenendael H, Bauer FW, Rijzewijk JJ: Growth fraction in epidermal skin disorders determined by the monoclonal antibody Ki 67. J Invest Dermatol 1987;89: 312–316.
19.
Bravo R, MacDonald-Bravo H: Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: Association with DNA replication sites. J Cell Biol 1987; 105:1549–1554.
20.
Hall PA, McKee PH, Du P, Menage H, Dover R, Lane DP: High levels of p53 protein in UV irradiated normal human skin. Oncogene 1993; 8:203–207.
21.
Wolff HH, Gnas W: Immunocytochemical detection of in vitro incorporated 5-bromodeoxyuridine in paraffin sections of human epidermis. Arch Dermatol Res 1989;281: 209–212.
22.
Baserga R, Malamud D: Autoradiography, Techniques and Applications. New York, Hoeber, 1969.
23.
Suurmeijer AJH, Boon ME: Optimizing keratin and vimentin retrieval in formalin fixed, paraffin embedded tissue with the use of heat and metal salts. Appl Immunohistochem 1993; 1:143–148.
24.
Boezeman JBM, Bauer FW, Degrood RM: Flow cytometric analysis of the recruitment of G0 cells in human epidermis in vivo following tape stripping. Cell Tissue Kinet 1987;20: 99–107.
25.
Potten CS, Morris RJ: Epithelial stem cells in vivo. J Cell Sci Suppl 1988;10:45–62.
26.
Dover R, Potten CS: Cell cycle kinetics of cultured human epidermal keratinocytes. J Invest Dermatol 1983;80:423–429.
27.
Heenen M, De Graef CH, Parent D, De Dobbeleer G: Renewal and differentiation of keratinocytes cultured on dead deepidermized dermis. Cell Prolif 1992;25:311–319.
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