Abstract
Purpose::
The secretion of the lacrimal gland provides 95% of the aqueous tears, which are essential for lubrication, nutrition and protection of the ocular surface. Long term studies of acinar lacrimal gland cells in-vitro are complicated by low proliferation rate and fast loss of cell function on plastic. Aim of this study was to analyze the effect of amniotic membrane as a carrier on the proliferation and secretory function of lacrimal gland acinar cells in a rabbit model.
Methods::
Lacrimal gland acinar cells from Chinchilla Bastard- and New Zealand White rabbits of different sexes were isolated and cultured on denuded amniotic membrane in modified HSM-medium. Cells were analysed by light- and electron-microscopy. Cell viability was analysed by means of the Calcein/AM-Assay and secretory response to carbachol was tested by measuring the ß-hexosaminidase activity.
Results::
By day 3 single cells and small groups of cells were found on the amniotic membrane. Cells showed polarity, with a basal nucleus and large electron lucent apical granules. Between day 7 and 14 the cell groups increased in size and acini with a central lumen were noted. On day 28 cellular organisation into acini, accumulation of secretory material in the lumen and desmosome formation connecting the apical cell structures was observed. However, between day 14 and 28 the number of cytoplasmatic granules decreased and fibroblast like cells appeared. The stimulation with carbachol showed an increase of ß-hexosaminidase release until day 14.
Conclusions::
Acinar lacrimal gland cells can be successfully cultured on amniotic membrane up to 28 days, with a secretory response to carbachol up to 14 days. This may be a suitable in-vitro model for studying acinar cell proliferation and cell function.