Engineering of a Secretory Active Three-Dimensional Lacrimal Gland Construct on the Basis of Decellularized Lacrimal Gland Tissue

Abstract

Lacrimal gland (LG) insufficiency is a main cause for severe dry eye leading to pain, visual impairment, and eventually loss of sight. Engineering of transplantable LG tissue with secretory capacity is a desirable goal. In this study, a three-dimensional decellularized LG (DC-LG) scaffold with preserved LG morphology was generated by treatment with 1% sodium deoxycholate and DNase solution using porcine LG tissue. To address clinical applicability, the primary in vitro culture of secretory active LG cells from a small tissue biopsy of 1.5 mm diameter was introduced and compared with an established isolation method by enzymatic digestion. Cells from both isolation methods depicted an epithelial phenotype, maintained their secretory capacity for up to 30 days, and exhibited progenitor cell capacity as measured by aldehyde dehydrogenase-1 activity, side population assay, and colony-forming units. Cells from passage 0 were reseeded into the DC-LG and secretory active cells migrated into the tissue. The cells resembled an LG-like morphology and the constructs showed secretory activity. These results demonstrate the possibility of engineering a secretory competent, three-dimensional LG construct using LG cells expanded from a small tissue biopsy and DC-LG as a matrix that provides the native structure and physiological niche for these cells.

Development of lacrimal gland spheroids for lacrimal gland tissue regeneration

“…This was performed as previously described (Spaniol et al, ). Briefly, lacrimal glands were harvested from 8‐week‐old pigs and cultures initiated from 1‐mm 3 explants on a growth‐arrested 3T3 feeder layer.…”

“…A control sample was then collected, and carbachol was added (final concentration: 100 μM) for a further 30 min before this was also collected. Quantification of β‐hexosaminidase activity was performed as described previously (Spaniol et al, ).…”

“…Epithelial acini are considered to be the functional units of the lacrimal gland since these produce lacrimal fluid (Kanski & Bowling, ; Lang, ). As a first step towards tissue engineering of lacrimal gland tissue, methods for culture of these lacrimal gland epithelial cells have been established (Spaniol et al, ; Tiwari et al, ). Others have progressed to evaluating various synthetic and nonsynthetic scaffolds for their ability to support lacrimal gland cell expansion whilst maintaining a functional phenotype (reviewed in Massie et al, ).…”

Lacrimal gland development: From signaling interactions to regenerative medicine

“…However, following a two week period, continuous cell death was frequently displayed (Schrader et al, 2009). The same group recently described a non-immunogenic decellularized tissue matrix derived from pig lacrimal gland, which allows lacrimal gland epithelial cells seeded on this scaffold to maintain acinar-like structures and secretory ability for up to 30 days (Spaniol et al, 2015). In a similar effort, rabbit lacrimal gland progenitor cells derived from a sphere-forming culture have been shown to differentiate and display secretory function in either a decellularized lacrimal gland matrix or 3-D collagen gel (Lin et al, 2016).…”

An In Vitro Model for the Ocular Surface and Tear Film System

“…Considering the ocular surface and tear film, researchers have been investigating the in vitro modeling of several components individually, including conjunctival epithelium 17 and lacrimal glands 18, 19 . In these models, cell morphology and phenotype resembled the equivalent cells in vivo .…”