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Development of lacrimal gland spheroids for lacrimal gland tissue regeneration

 
: Massie, Isobel; Spaniol, Kristina; Barbian, Andreas; Geerling, Gerd; Metzger, Marco; Schrader, Stefan

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Journal of tissue engineering and regenerative medicine 12 (2018), No.4, pp.e2001-e2009
ISSN: 1932-6254
ISSN: 1932-7005
English
Journal Article
Fraunhofer IGB ()
cytokeratin; endoglin; Hermes antigen; lysozyme

Abstract
Severe dry eye syndrome resulting from lacrimal gland (LG) dysfunction can cause blindness, yet treatments remain palliative. In vitro reconstruction of LG tissue could provide a curative treatment. We aimed to combine epithelial cells with endothelial cells and mesenchymal stem cells (MSCs) to form a 3D functional unit. Epithelial cells and MSCs were isolated from porcine LG; endothelial cells were isolated from human foreskin. MSCs were characterised (flow cytometry and differentiation potential assays). All 3 cell types were combined on Matrigel and spheroid formation observed. Spheroids were characterised [immunohistochemistry (IHC) and transmission electron microscopy] and function assessed (β-hexosaminidase assay). Spheroids were transferred to decellularised jejunum (SIS-Muc) in dynamic cultures for 1 week before further characterisation. MSCs did not express CD31 but expressed CD44 and CD105 and differentiated towards osteogenic and adipogenic lineages. Spheroids formed on Matrigel within 18 hr, contracting to ~10% of the well area (p .005). IHC revealed presence of all 3 cells within spheroids. Transmission electron microscopy revealed cell–cell contacts and polarisation at the apical surface. In static cultures, function was increased in spheroids cf. monolayer controls (p <.05) but over 72 hr, spheroid function (p <.05), viability (p <.05), and proliferation decreased, whilst apoptosis increased. On SIS-Muc under dynamic culture, however, spheroids continued to proliferate to repopulate SIS-Muc. IHC revealed LG epithelial cells coexpressing pan-cytokeratin and lysozyme, as well as endothelial cells and MSCs and cells remained capable of responding to carbachol (p <.05). These spheroids could form the basis of a regenerative medicine treatment approach for dry eye syndrome. In vivo studies are required to evaluate this further.

: http://publica.fraunhofer.de/documents/N-512441.html