Amniotic membrane as a carrier for lacrimal gland acinar cells

Schrader, Stefan; Wedel, Thilo; Kremling, C.; Laqua, H.; Geerling, Gerd

doi:10.1007/s00417-007-0612-7

Cited by 24 publications

(16 citation statements)

References 20 publications

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“…After 3 days of culture on amniotic membrane, we found strong β-hexosaminidase secretion which was about three to four times that of the baseline. The secretory resonse decreased to approximately twice that of the baseline at day 7 and was substantially diminished beyond day 7 2. Similar observations were made by Andersson et al , who reported optimal secretory response of acinar cells after 2–3 days in culture,17 and Schechter et al , who noted a strong reduction in acinar cell secretion beyond day 7 5.…”

Section: Discussionsupporting

confidence: 84%

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Cultivation of lacrimal gland acinar cells in a microgravity environment

Schrader

Kremling

Klinger

et al. 2009

British Journal of Ophthalmology

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Background: A Rotary Cell Culture System (RCCS) allows the creation of a microgravity environment of low shear force, high-mass transfer, and 3-dimensional cell culture of various cell types. Aim of this study was to evaluate the growth pattern and the secretory function of rabbit lacrimal gland acinar cells in a microgravity environment using a RCCS. Methods:Lacrimal gland acinar cells from male New Zealand White rabbits were isolated and cultured in a RCCS up to 28 days. Cells were analysed by light and electron microscopy and apoptosis was assessed by the TUNEL-Assay at day 7, 14, 21 and 28. Secretory function was tested by measuring the ß-hexosaminidase activity.Results: After 7 days of culture, spheroidal aggregates were found inside the RCCS. The spheroids consisted of acinus-like cell conglomerates. Apoptotic centers inside the spheroids were observed at all time points by means of the TUNEL-Assay. Evaluation of the secretory function revealed ß-hexosaminidase release after carbachol stimulation which decreased over the culture period. Conclusion:A simulated microgravity environment promotes the development of three dimensional cell spheroids containing viable acinar cells up to 28 days. Due to the evolving central apoptosis, it is unlikely that such simple three dimensional cell communities can serve as tissue equivalents for clinical transplantation, but they promise opportunities for further applications in basic and applied cell research on lacrimal gland cells.

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Section: Discussionsupporting

confidence: 84%

“…Several methods have been developed to maintain lacrimal gland cells in vitro,2 3 4 5 6 but preservation of the acinar phenotype and secretory function has proven to be difficult. Rotary cell-culture systems (RCCS) were originally designed to predict the impact of the microgravity environment in space for the culture of cells 7.…”

mentioning

confidence: 99%

Cultivation of lacrimal gland acinar cells in a microgravity environment

Schrader

Kremling

Klinger

et al. 2009

British Journal of Ophthalmology

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“…The simplified procedure did not compromise the purity of LGAC cells and helped to increase the longevity of the primary culture, compatible with recently published protocols (10,11) . Our hypothesis is that reduced manipulation during isolation contributes markedly to improving and extending cell viability and behavior during culture.…”

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confidence: 66%

“…Several attempts to enhance the growth of acinar cells in culture have been made, but after a span of approximately 3 weeks, LGACs exhibited decreased viability, with high numbers of apoptotic cells (11)(12)(13)(14) . Exocrine acinar cells are fragile, post-mitotic epithelial cells with marked polarity.…”

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confidence: 99%

Lacrimal gland primary acinar cell culture: the role of insulin

Malki

Dias

Jorge

et al. 2016

Arquivos Brasileiros De Oftalmologia

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The condition of lacrimal glands (LG) is influenced by components of the immune, neural, and endocrine systems (1)(2)(3)(4)(5) . Insulin is thought to be of major importance, based on observations related to its deprivation in diabetes mellitus (DM), on LG, tear film, and ocular surface in humans, animal models, and in culture (6)(7)(8) . Lacrimal gland acinar cell (LGAC) culture is an appropriate model for improving the understanding of the influence of insulin on these cells.In previous studies, the culture of LGACs from different species was performed over an average period of 21 days (9,10) . Several attempts to enhance the growth of acinar cells in culture have been made, but after a span of approximately 3 weeks, LGACs exhibited decreased viability, with high numbers of apoptotic cells (11)(12)(13)(14) . Exocrine acinar cells are fragile, post-mitotic epithelial cells with marked polarity. This indicates that the apical and basal sides are clearly positioned but also that intracellular organelles are located at one or the other pole, depending on their cellular functions. Thus, an adequate extracellular matrix and other specific ingredients are needed in the culture media to mimic in vivo conditions (15,16) . The handling during isolation and culture procedure of LGACs should be gentle ABSTRACT Purpose: The goal of the present study was to establish a protocol for primary culture of lacrimal gland acinar cells (LGACs) and to assess the effect of adding insulin to the culture media. Methods:LGACs were isolated and cultured from lacrimal glands of Wistar male rats. The study outcomes included cell number, viability, and peroxidase release over time and in response to three concentrations of insulin (0.5, 5.0, and 50.0 μg/mL). Results: In LGAC primary culture, cells started to form clusters by day 3. There was a time-response pattern of peroxidase release, which rose by day 6, in response to carbachol. Culture viability lasted for 12 days. An insulin concentration of 5.0 μg/mL in the culture medium resulted in higher viability and secretory capacity. Conclusions:The present method simplifies the isolation and culture of LGACs. The data confirmed the relevance of adding insulin to maintain LGACs in culture. Keywords

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“…It also evident that chondrogenic differentiation were enhanced following chemical induction as compared to monolayer cultures, making HAM a potentially suitable material to enhance cellular phenotypic expression. In other studies, the role of HAM, when seeded with stem cells for MSC differentiation, has been documented although all of it were performed in in vivo transplantation without prior in vitro differentiation (Schrader et al 2007;Yang et al 2008). The rationale for phenotypic differentiation was based on the notion that environment induction was adequate to promote the changes without the need for artificial stimulation.…”

Section: Discussionmentioning

confidence: 95%

Human amnion as a novel cell delivery vehicle for chondrogenic mesenchymal stem cells

et al. 2009

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This study investigates the feasibility of processed human amnion (HAM) as a substrate for chondrogenic differentiation of mesenchymal stem cells (MSCs). HAM preparations processed by air drying (AD) and freeze drying (FD) underwent histological examination and MSC seeding in chondrogenic medium for 15 days. Monolayer cultures were used as control for chondrogenic differentiation and HAMs without cell seeding were used as negative control. Qualitative observations were made using scanning electron microscopy analysis and quantitative analyses were based on the sulfated glycosaminoglycans (GAG) assays performed on day 1 and day 15. Histological examination of HAM substrates before seeding revealed a smooth surface in AD substrates, while the FD substrates exhibited a porous surface. Cell attachment to AD and FD substrates on day 15 was qualitatively comparable. GAG were significantly highly expressed in cells seeded on FD HAM substrates. This study indicates that processed HAM is a potentially valuable material as a cell-carrier for MSC differentiation.

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Amniotic membrane as a carrier for lacrimal gland acinar cells

Abstract: Acinar lacrimal gland cells can be successfully cultured on amniotic membrane up to 28 days, with a secretory response to carbachol up to 21 days. This model may be used for further experimental work, to elucidate cellular mechanisms in normal and diseased lacrimal tissue.

Cited by 24 publications

References 20 publications

Cultivation of lacrimal gland acinar cells in a microgravity environment

Cultivation of lacrimal gland acinar cells in a microgravity environment

Lacrimal gland primary acinar cell culture: the role of insulin

Human amnion as a novel cell delivery vehicle for chondrogenic mesenchymal stem cells

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