Culture of rabbit meibomian gland using collagen gel

Rabbit whole meibomian glands were isolated by microsurgical dissection and enzymatic digestion. Both ductal and acinar elements could be separated by subsequent microsurgical manipulation. Epithelial outgrowth could be derived from either the whole gland or the ductal/acinar elements using collagen gel in the serum-containing, growth factor-enriched medium. Two types of outgrowths were observed: a discontinuous spindle-shaped epithelial cell monolayer in which oil-red-O (ORO)-stained meibum containing single cells and cell aggregates were evenly distributed, and a continuous cuboidal epithelial monolayer in which oil-red-O-stained cells formed a gradient with single cells and cell aggregates preferentially distributed in the periphery of the outgrowth. Thus, meibum production and meibomian gland differentiation can be maintained in collagen gel culture and ductal elements may contain progenitor cells for acinar differentiation. This culture system will allow future exploration of the regulatory mechanism of meibomian gland differentiation at the cellular level.