Use of primary keratinocyte cultures from plucked human hairs for analysis of gap junctional intercellular communication.
Hair follicles are a convenient source of human keratinocytes that are easily derived by non-invasive means. In order to test whether such cells could be used for gap junctional intercellular communication (GJIC) studies for the assessment of tumour-promoting activity of environmental chemicals, primary cultures were initiated from plucked hairs of various donors. GJIC measurements were performed when colonies reached a size of 5 mm in diameter, by microinjection of the fluorescent dye Lucifer Yellow CH into single cells at the colony periphery and scoring dye transfer to surrounding cells. The GJIC of cells that had begun to differentiate (appearance of cornified envelopes) can be quantitated more easily by switching cells to a low calcium (0.05-mm) medium. The tumour promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) inhibited GJIC in a concentration- and time-dependent manner, resembling the TPA response of mouse primary epidermal keratinocytes. Thus, hair follicle cultures can be readily used for GJIC studies and should be appropriate for other in vitro assays requiring attached cells.
Swierenga
S H
SH
Drugs Directorate, Health and Welfare Canada, Banting Building, Tunney's Pasture, Ottawa, Ontario K1A OL2, Canada.
Fitzgerald
J
J
Yamasaki
H
H
Piccoli
C
C
Goldberg
M
M