Isolation of human sebaceous glands and cultivation of sebaceous gland-derived cells as an in vitro model

Abstract

An experimental technique is presented as an in vitro model for the study of human sebaceous gland-derived cells. Intact sebaceous glands were isolated from full-thickness human skin after incubation in dispase (2.4 U/ml) and in deoxyribonuclease (0.02%) by using microsurgical instruments under microscopical observation of the epidermal underface. Subsequently, the ducts of the glands were removed, the isolated gland lobules were seeded on a 3T3-cell feeder layer in Dulbecco's modified Eagle's medium and Ham's F 12 medium (3:1) supplemented with fetal calf serum (10%), L-glutamine, antibiotics, epidermal growth factor (10 ng/ml), hydrocortisone (0.4 microgram/ml), and cholera toxin (10(-9) M), and were then cultivated in a CO2-incubator at 37 degrees C. After 2-3 wk cell outgrowths resulting from the periphery of the gland lobules were obtained and dispersed cells were passaged for three subcultures with or without 3T3-cell feeder layer. The cultured cells preserved in vitro morphologic characteristics and differentiation patterns comparable to those described for normal human sebocytes in vivo, with a high rate of viable cells. Their labeling pattern with MoAb showed close similarities to the pattern reported for sebocytes in vivo but differences to the pattern of keratinocytes in vivo and in vitro. In their cytoplasm oil red and nile red stained droplets were detected, and the observed density and distribution evidenced in vitro lipogenesis. The technique presented here may provide a promising model for further experimental studies on sebaceous gland cell development and function.

Dermal cysts of the rhino mouse develop into unopened sebaceous glands

“…In addition, dermal cysts express a 47-kDa keratin, that is typical of basal cells of murine stratified epithelia (Schweizer 1993). This said, our results confirmed the presence of keratin 14 and are in agreement with those obtained for the human pilosebaceous unit (Kurokawa et al 1988;Xia et al 1989). There is evidence that they may represent hard α-keratins, but this would be surprising since the expression of these keratins is invariably associated with the formation of hard keratinized structures.…”

In Vitro Models for the Evaluation of Anti-acne Technologies

“…The in vivo data appeared consistent with early in vitro experiments that failed to yield viable sebocytes during extended culture conditions (27)(28)(29). This resulted in a similar but less stringent need as organ culture for ample biopsies to supply sufficient sebocytes for experimentation.…”

SACK-Expanded Hair Follicle Stem Cells Display Asymmetric Nuclear Lgr5 Expression With Non-Random Sister Chromatid Segregation

“…6D) that stained with oil-red (Fig. 6E), indicative of sebocyte differentiation with production of sebum30. In addition, transmission electron microscopy analysis of cells maintained under these conditions revealed a high frequency of fully formed primary cilia, a differentiated cell feature that is unusual for many types of cultured cells (Fig.…”