title: Cryopreservation and hypothermic storage of lacrimal gland: towards enabling delivery of regenerative medicine therapies for treatment of dry eye syndrome.
abstract: Severe dry eye syndrome (DES) can cause painful loss of vision and may result from lacrimal gland dysfunction. Current treatments are palliative, so a causative therapy is desirable. The ability to (cryo)preserve lacrimal gland tissue or epithelial cells would simplify this. Here, lacrimal gland tissue was cryopreserved in 10% dimethylsulphoxide in liquid nitrogen, or stored at 4 °C in culture medium for up to 7 days, and compared with fresh tissue using immunohistochemistry. Cultures were initiated from fresh and stored tissue, and cells characterised in P1 for proliferation (WST-1), colony-forming efficiency (CFE) and secretory capacity (immunocytochemistry and β-hexosaminidase activity assay). Tissue stored for > 3 days at 4 °C displayed grossly altered tissue architecture when compared with fresh tissue, decreased acinus density and increased caspase-3 activity. Cryopreserved tissue showed less obvious signs of damage without caspase-3 activation. Storage at 4 °C and cryopreservation delayed epithelial outgrowth compared with that from fresh tissue initially (p  <  0.05) but, by day 9, all explants showed comparable outgrowth (~90%), except tissue stored at 4 °C for 3 or 7 days (p  <  0.05 compared with fresh tissue). Epithelial cell yields per explant were similar from fresh and stored tissue, apart from tissue stored at 4 °C for 7 days (p  <  0.01). In P1, epithelial cells from fresh and stored tissue were largely equivalent in terms of: proliferation; CFE (~21%); Rab3D, HexA and lysozyme expression; mucin production; and β-hexosaminidase activity. These data demonstrate that cryo(preservation) of lacrimal gland tissue and cells is possible, which may enable use of autologous cells in regenerative medicine approaches to treating DES. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Labor für Experimentelle Ophthalmologie, Universitätsklinikum Düsseldorf, Life Science Center, Düsseldorf, Germany.
authors.forename: I
authors.initials: I
authors.lastname: Massie
authors.affiliation: Augenklinik, Universitätsklinikum Düsseldorf, Düsseldorf, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Spaniol
authors.affiliation: Augenklinik, Universitätsklinikum Düsseldorf, Düsseldorf, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Geerling
authors.affiliation: Augenklinik, Universitätsklinikum Düsseldorf, Düsseldorf, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Schrader
mesh_terms: D016475:3T3 Cells; D000818:Animals; D020169:Caspases / Q000378:metabolism; D049109:Cell Proliferation; D002470:Cell Survival; D003080:Cold Temperature; D003114:Colony-Forming Units Assay; D015925:Cryopreservation; D015352:Dry Eye Syndromes / Q000628:therapy*; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000166:cytology; D007765:Lacrimal Apparatus / Q000637:transplantation*; D051379:Mice; D009077:Mucins / Q000378:metabolism; D044968:Regenerative Medicine / Q000379:methods*; D034421:Sus scrofa; D013997:Time Factors
chemical_list: D009077:Mucins; D020169:Caspases
keywords: cryopreservation; dry eye syndrome; hypothermic storage; lacrimal gland; regeneration; tissue engineering