Isolation and Propagation of Mesenchymal Stem Cells from the Lacrimal Gland

You, Samantha; Kublin, Claire L.; Avidan, Orna; Miyasaki, David; Zoukhri, Driss

doi:10.1167/iovs.10-5686

Cited by 60 publications

(63 citation statements)

References 36 publications

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“…Previous studies have shown that mesenchymal stem cells exist in both lacrimal glands [57] and pancreas, specifically in ductal regions [58]. Thus, it is possible that loss of P2X7 enhances the differentiation of mesenchymal progenitor cells toward the adipocyte lineage, resulting in increased adipocyte numbers in periductal regions of the pancreas and lacrimal glands.…”

Section: Discussionmentioning

confidence: 99%

Loss of P2X7 nucleotide receptor function leads to abnormal fat distribution in mice

Beaucage

Xiao

Pollmann

et al. 2013

Purinergic Signalling

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The P2X7 receptor is an ATP-gated cation channel expressed by a number of cell types. We have shown previously that disruption of P2X7 receptor function results in downregulation of osteogenic markers and upregulation of adipogenic markers in calvarial cell cultures. In the present study, we assessed whether loss of P2X7 receptor function results in changes to adipocyte distribution and lipid accumulation in vivo. Male P2X7 loss-of-function (KO) mice exhibited significantly greater body weight and epididymal fat pad mass than wild-type (WT) mice at 9 months of age. Fat pad adipocytes did not differ in size, consistent with adipocyte hyperplasia rather than hypertrophy. Histological examination revealed ectopic lipid accumulation in the form of adipocytes and/or lipid droplets in several non-adipose tissues of older male KO mice (9-12 months of age). Ectopic lipid was observed in kidney, extraorbital lacrimal gland and pancreas, but not in liver, heart or skeletal muscle. Specifically, lacrimal gland and pancreas from 12-month-old male KO mice had greater numbers of adipocytes in perivascular, periductal and acinar regions. As well, lipid droplets accumulated in the renal tubular epithelium and lacrimal acinar cells. Blood plasma analyses revealed diminished total cholesterol levels in 9-and 12-month-old male KO mice compared with WT controls. Interestingly, no differences were observed in female mice. Moreover, there were no significant differences in food consumption between male KO and WT mice. Taken together, these data establish novel in vivo roles for the P2X7 receptor in regulating adipogenesis and lipid metabolism in an age-and sex-dependent manner.

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Section: Discussionmentioning

confidence: 99%

Loss of P2X7 nucleotide receptor function leads to abnormal fat distribution in mice

Beaucage

Xiao

Pollmann

et al. 2013

Purinergic Signalling

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“…Recently, You et al isolated the progenitor cells from the injured murine LG and identified them as mesenchymal stem cells. 16 They were not able to isolate progenitor cells from the uninjured murine LG. 16 We recently developed a method to culture MECs from uninjured, adult rat LGs.…”

mentioning

confidence: 99%

“…16 They were not able to isolate progenitor cells from the uninjured murine LG. 16 We recently developed a method to culture MECs from uninjured, adult rat LGs. 5 MECs typically take 3-4 weeks to differentiate.…”

mentioning

confidence: 99%

Isolation and Characterization of Progenitor Cells in Uninjured, Adult Rat Lacrimal Gland

Shatos

Haugaard‐Kedström²,

Hodges³

et al. 2012

Invest. Ophthalmol. Vis. Sci.

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“…Fibroblasts appeared by the third week of culture. None of these obser vations had been registered in our previous experiments or reported by other authors in studies using shorter culture periods (10,17,18,21) . The use of a combination of quinolone (ofloxacin) and aminoglycoside (gentamicin) was effective in terms of preventing contamination of the culture.…”

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confidence: 63%

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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Isolation and Propagation of Mesenchymal Stem Cells from the Lacrimal Gland

Abstract: Murine lacrimal glands contain mesenchymal stem cells that seem to play a pivotal role in tissue repair.

Cited by 60 publications

References 36 publications

Loss of P2X7 nucleotide receptor function leads to abnormal fat distribution in mice

Loss of P2X7 nucleotide receptor function leads to abnormal fat distribution in mice

Isolation and Characterization of Progenitor Cells in Uninjured, Adult Rat Lacrimal Gland

Lacrimal gland primary acinar cell culture: the role of insulin

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