[Outcomes of 3D culture of human lacrimal gland cells (pilot study)].

Abstract

Objective: The study tested a method for 3D culture of human lacrimal gland (LG) cells with the formation of structures suitable for subsequent investigation of the pathogenetic mechanisms underlying its diseases.

Material and methods: Material for microbiological culture was obtained during a diagnostic biopsy performed as part of surgical correction of recurrent LG prolapse. No morphological changes were detected in the LG tissue. Cells were isolated from the tissue specimen by sequential mechanical and enzymatic dissociation. The resulting cell populations were separated by differential trypsinization. Spheroids were formed by embedding the cell suspension in a dome of concentrated Matrigel solution. Cell viability was assessed using double staining with calcein-AM and ethidium homodimer, and expression of specific markers (Muc5AC, Na/K-ATPase, panCK) was analyzed by immunocytochemistry.

Results: A sufficient number of viable cells was cultured from the postoperative material to form spheroids in Matrigel. Spheroids derived from epithelial and fibroblast-like human LG cells were obtained and characterized. Epithelial cells expressed Muc5AC and Na/K-ATPase, while fibroblasts formed characteristic reticular structures in the matrix.

Conclusion: The tested protocol allows for the formation of viable spheroid structures that retain the morphofunctional properties of native LG tissue. The developed in vitro model of the human LG can be used to study the pathogenetic mechanisms of its diseases, particularly chronic nonspecific dacryoadenitis.