The effect of choleragen and epidermal growth factor on proliferation and maturation in vitro of human ectocervical cells |
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Authors: | Margaret A Stanley Kirsten Dahlenburg |
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Institution: | (1) Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QP, U.K., England;(2) Department of Pathology, University of Adelaide, Tennis Court Road, Cambridge, CB2 1QP, U.K., South Australia;(3) Present address: Institute of Medical and Veterinary Sciences, Adelaide, South Australia |
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Abstract: | Summary The role of choleragen (CT) and epidermal growth factor (EGF) has been examined in relation to the control of growth and differentiation
of adult human cervical epithelial (HCE) cells derived from the ectocervix. Cervical biopsies derived from hysterectomy specimens
were trypsin disaggregated and HCE cells were plated at 5×103/cm2 in the presence of 2×104/cm2 lethally irradiated Swiss 3T3 fibroblasts. Cultures were grown in Liebovitz medium supplemented with 10% fetal bovine serum
and hydrocortisone. Epidermal growth factor at 10 ng/ml and choleragen at 10−10
M were added to cultures either singly or in combination. DNA replication in these cultures was measured autoradiographically
after exposing cells to tritiated thymidine for 2 h. Differentiation was assessed histochemically by determining glycogen
accumulation using the periodic acid Schiff technique. Choleragen increased colony plating efficiency by at least a factor
of two but had no effect on colony size Epidermal growth factor did not increase plating efficiency but did increase colony
size. In EGF treated colonies DNA replication occurred throughout the colony compared to CT treated colonies in which replication
was restricted to the periphery. In the absence of EGF, population doublings achieved in culture did not exceed 32 and glycogen
accumulation was evident in cells early in culture life. Colonies treated with EGF exhibited glycogen accumulation late in
culture life and the EGF treated cells achieved at least 50 population doublings in culture. The results are discussed in
relation to the role of EGF and choleragen on cell differentiation. |
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Keywords: | cervical cell cultures choleragen EGF differentiation |
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