Flow sorting from organ material by intracellular markers

Moerch, Ulrik; Nielsen, Henriette Svarre; Lundsgaard, Dorthe; Oleksiewicz, Martin B.

doi:10.1002/cyto.a.20418

Cited by 12 publications

(10 citation statements)

References 18 publications

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“…Additional enrichment of C-cells before cell culture by, e.g. FACS-sorting, as reported by Moerch et al (2007), is hindered by the lack of an established cell surface marker for C-cells. Despite some current limitations, the cell culture method and results reported here may open new avenues to study the putative physiological and/or pathological roles of GLP1 and GLP1R agonists on normal, non-transformed primary C-cells.…”

Section: Figurementioning

confidence: 99%

Effect of GLP1R agonists taspoglutide and liraglutide on primary thyroid C-cells from rodent and man

Boess¹,

Bertinetti-Lapatki²,

Zoffmann³

et al. 2013

Journal of Molecular Endocrinology

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Glucagon-like peptide 1 (GLP1) analogs have been associated with an increased incidence of thyroid C-cell hyperplasia and tumors in rodents. This effect may be due to a GLP1 receptor (GLP1R)-dependent mechanism. As the expression of GLP1R is much lower in primates than in rodents, the described C-cell proliferative lesions may not be relevant to man. Here, we aimed to establish primary thyroid cell cultures of rat and human to evaluate the expression and function of GLP1R in C-cells. In our experiments, GLP1R expression was observed in primary rat C-cells (in situ hybridization) but was not detected in primary human C-cells (mRNA and protein levels). The functional response of the cultures to the stimulation with GLP1R agonists is an indirect measure of the presence of functional receptor. Liraglutide and taspoglutide elicited a modest increase in calcitonin release and in calcitonin expression in rat primary thyroid cultures. Contrarily, no functional response to GLP1R agonists was observed in human thyroid cultures, despite the presence of few calcitonin-positive C-cells. Thus, the lack of functional response of the human cultures adds to the weight of evidence indicating that healthy human C-cells have very low levels or completely lack GLP1R. In summary, our results support the hypothesis that the GLP1R agonist-induced C-cell responses in rodents may not be relevant to primates. In addition, the established cell culture method represents a useful tool to study the physiological and/or pathological roles of GLP1 and GLP1R agonists on normal, non-transformed primary C-cells from rats and man.

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Section: Figurementioning

confidence: 99%

Effect of GLP1R agonists taspoglutide and liraglutide on primary thyroid C-cells from rodent and man

Boess¹,

Bertinetti-Lapatki²,

Zoffmann³

et al. 2013

Journal of Molecular Endocrinology

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“…20 Tween 20 is usually used at a concentration of 0.05-0.1% for lysing cells. 21,22 Triton X-100 at a concentration of 0.05-1.0% enables cell permeabilization for immunostaining, 21,23,24 and SDS at 0.3-2% is used for cell homogenates. 21,23 In this study, in order to enable penetration of MB solution into the interstices of fractured lines or cracks, the use of these three detergents at concentrations of 0.1% or 1.0% were examined.…”

Section: Discussionmentioning

confidence: 99%

Basic study of the use of laser on detection of vertical root fracture

Kimura

Tanabe

Amano

et al. 2009

Journal of Dentistry

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“…FACS identification and/or sorting of several structures presenting a size similar to the secretory granules (subcellular structures, bacteria) using some of the above-mentioned alternate labeling approaches were reported. [18, 36–38]). Doing so, the user can virtually sort and actively exclude any event in a defined window which, in turn, leads to much increased purity of the sorted material.…”

Section: Discussionmentioning

confidence: 99%

Flow cytometry‐assisted purification and proteomic analysis of the corticotropes dense‐core secretory granules

et al. 2008

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The field of organellar proteomics has emerged as an attempt to minimize the complexity of the proteomics data obtained from whole cell and tissue extracts while maximizing the resolution on the protein composition of a single subcellular compartment. Standard methods involve lengthy density-based gradient and/or immunoaffinity purification steps followed by extraction, onedimensional or two-dimensional gel electrophoresis, gel staining, in-gel tryptic digestion and protein identification by mass spectrometry. In this paper, we present an alternate approach to purify subcellular organelles containing a fluorescent reporter molecule. The gel-free procedure involves fluorescence-assisted sorting of the secretory granules followed by gentle extraction in a buffer compatible with tryptic digestion and mass-spectrometry. Once the subcellular organelle labeled, this procedure can be done in a single day, requires no major modification to any instrumentation and can be readily adapted to the study of other organelles. When applied to corticotrope secretory granules, it led to a much enriched granular fraction from which numerous proteins could be identified through mass spectrometry.

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Flow sorting from organ material by intracellular markers

Cited by 12 publications

References 18 publications

Effect of GLP1R agonists taspoglutide and liraglutide on primary thyroid C-cells from rodent and man

Effect of GLP1R agonists taspoglutide and liraglutide on primary thyroid C-cells from rodent and man

Basic study of the use of laser on detection of vertical root fracture

Flow cytometry‐assisted purification and proteomic analysis of the corticotropes dense‐core secretory granules

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