Two types of normal human breast epithelial cells derived from reduction mammoplasty: phenotypic characterization and response to SV40 transfection

Abstract

A culture method to grow two morphologically distinguishable normal human breast epithelial cell types derived from reduction mammoplasty has been developed. Type I cells were characterized by a more variable cell shape, smooth cell colony boundaries, the expression of epithelial membrane antigen (EMA) and keratin 18 and the non-expression of keratin 14 and alpha 6 integrin. In addition, the Type I cells were growth stimulated by fetal bovine serum (FBS) and were deficient in gap junctional intercellular communication (GJIC). In contrast, Type II cells were characterized by a uniform cell shape, expression of keratin 14 and alpha 6 integrin and the non-expression of EMA and keratin 18. In addition, Type II cells were growth inhibited by FBS and were proficient in GJIC. Type I cells can be induced by cholera toxin to change their morphology to a Type II cell morphology. Hence, Type I cells antigenically resemble luminal epithelial cells, while the Type II cells more closely resemble basal epithelial cells. Type I and Type II cells were transfected with SV40 DNA. Clones with extended lifespans were obtained from both Type I and Type II cells by SV40 transfection. Some (2/9) of the SV40-transfected Type I cell clones became immortal (> 100 cumulative population doubling level), whereas none (0/8) of the SV40-transfected Type II cell clones became immortal. The SV-40-transfected Type I and Type II cell-derived extended life clones and immortal cell lines phenotypically resembled their parental cells with respect to EMA, keratin 14 and keratin 18 expression and GJIC. Each (9/9) of the SV40 transfected Type I cell clones grew in soft agar; none (0/8) of the SV40-transfected Type II cell clones were capable of growing in soft agar. These results provide evidence that normal human breast epithelial cells, derived from reduction mammoplasty, can be separated into two morphologically and antigenically different cell types and that these two different cell types significantly differ in their response to an oncogenic (SV40) stimulus.