Isolation of Mature (Peritoneum-Derived) Mast Cells and Immature (Bone Marrow-Derived) Mast Cell Precursors from Mice

Meurer, Steffen K.; Neß, M; Weiskirchen, Sabine; Kim, Philipp; Tag, Carmen G.; Kauffmann, Marlies; Huber, Michael; Weiskirchen, Ralf

doi:10.1371/journal.pone.0158104

Cited by 56 publications

(57 citation statements)

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“…Purified mast cells were transferred to 15% FBS and 5 µg/mL streptomycin and 50 U penicillin fortified RPMI-1640 for multiplication of mast cells. 47 Media was changed after every 3 days, and the culture was maintained up to 2 months.…”

Section: +mentioning

confidence: 99%

Inhibitory effect of a new orally active cedrol-loaded nanostructured lipid carrier on compound 48/80-induced mast cell degranulation and anaphylactic shock in mice

Chakraborty¹,

Kar²,

Kumari³

et al. 2017

IJN

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Background Type I hypersensitivity is an allergic reaction characterized by the overactivity of the immune system provoked by normally harmless substances. Glucocorticoids, anti-histamines, or mast cell stabilizers are the choices of treatment for type I hypersensitivity. Even though these drugs have the anti-allergic effect, they can have several side effects in prolong use. Cedrol is the main bioactive compound of Cedrus atlantica with anti-tumor, anti-oxidative, and platelet-activating factor inhibiting properties. Methods In this study, the preparation and anti-anaphylactic effect of cedrol-loaded nanostructured lipid carriers (NLCs) were evaluated. NLCs were prepared using Compritol ® 888 ATO and triolein as lipid phase and vitamin E d -α-tocopherylpolyethyleneglycol 1000 succinate, soya lecithin, and sodium deoxycholate as nanoparticle stabilizers. Results The average diameter of cedrol-NLCs (CR-NLCs) was 71.2 nm (NLC-C 1 ) and 91.93 nm (NLC-C 2 ). The particle had negative zeta potential values of −31.9 mV (NLC-C 1 ) and −44.5 mV (NLC-C 2 ). Type I anaphylactoid reaction in the animal model is significantly reduced by cedrol and cedrol-NLC. This in vivo activity of cedrol resulted that cedrol suppressed compound 48/80-induced peritoneal mast cell degranulation and histamine release from mast cells. Furthermore, compound 48/80-evoked Ca 2+ uptake into mast cells was reduced in a dose-dependent manner by cedrol and cedrol-NLC. Studies confirmed that the inhibition of type I anaphylactoid response in vivo in mice and compound 48/80-induced mast cell activation in vitro are greatly enhanced by the loading of cedrol into the NLCs. The safety of cedrol and CR-NLC was evaluated as selectivity index (SI) with prednisolone and cromolyn sodium as positive control. SI of CR-NLC-C 2 was found to be 11.5-fold greater than both prednisolone and cromolyn sodium. Conclusion Administration of CR-NLC 24 hours before the onset of anaphylaxis can prevent an anaphylactoid reaction. NLCs could be a promising vehicle for the oral delivery of cedrol to protect anaphylactic reactions.

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Section: +mentioning

confidence: 99%

Inhibitory effect of a new orally active cedrol-loaded nanostructured lipid carrier on compound 48/80-induced mast cell degranulation and anaphylactic shock in mice

Chakraborty¹,

Kar²,

Kumari³

et al. 2017

IJN

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“…Isolation and culture of peritoneal cell derived mast cells was performed as previous described protocol [22] with minor modifications. 5 ml of PBS and 2 ml of air was injected into peritoneal cavity of 14 weeks old mice.…”

Section: Methodsmentioning

confidence: 99%

Functional analysis of KIT gene structural mutations causing porcine dominant white phenotype by using genome edited mouse models

Sun

Qin

et al. 2019

Preprint

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24Dominant white phenotype in pigs is considered to be caused by two structural 25 mutations in KIT gene, including a 450-kb duplication encompassing the entire KIT 26 gene, and a splice mutation (G > A) at the first base in intron 17, which leads to the 27 deletion of exon 17 in mature KIT mRNA, and the production of KIT protein lacking a 28 critical catalytic domain of kinase. However, this speculation has not yet been validated 29 by functional studies. Here, by using CRISPR/Cas9 technology, we created two mouse 30 models mimicing the structural mutations of KIT gene in dominant white pigs, 31 including the splice mutation mouse model KIT D17/+ with exon 17 of one allele of KIT 32 gene deleted, and duplication mutation mouse model KIT Dup/+ with one allele of KIT 33 gene coding sequence (CDS) duplicated. We found that each mutation individually can 34 not cause dominant white phenotype. Splice mutation homozygote is lethal and 35 heterozygous mice present piebald coat. Inconsistent with previous speculation, we 36 found KIT gene duplication mutation did not confer the patched phenotype, and had no 37 obvious impact on coat color. Interestingly, combination of these two mutations lead to 38 dominant white phenotype. Further molecular analysis revealed that combination of 39 these two structural mutations could inhibit the kinase activity of the KIT protein, thus 40 reduce the phosphorylation level of PI3K and MAPK pathway associated proteins, 41 which may be related to the observed impaired migration of melanoblasts during 42 embryonic development, and eventually lead to dominant white phenotype. Our study 43 provides a further insight into the underlying genetic mechanisms of porcine dominant 44 white coat colour.45Author summary 46 KIT plays a critical role in control of coat colour in mammals. Two mutation 47 coexistence in KIT are considered to be the cause of the Dominant white phenotype in 48 pigs. One mutation is a 450-kb large duplication encompassing the entire KIT gene, 49 another mutation is a splice mutation causing the skipping of KIT exon 17. The 50 mechanism of these two mutations of KIT on coat color formation has not yet been 51 validated. In this study, by using genome edited mouse models, we found each 52 structural mutation individual does not lead dominant white phenotype, but 53 combination of these two mutations could lead to a nearly complete white coat colour 54 similar to pig dominant white phenotype, possibly due to the inhibition of the kinase 55 activity of the KIT protein, thus its signalling function on PI3K and MAPK pathways, 56 leading to impaired migration of melanoblasts during embryonic development, and 57 eventually lead to dominant white phenotype. Our study provides a further insight into 58 the underlying genetic mechanisms of porcine dominant white coat colour. 59 60 Introduction 61 Due to domestication and long term selection, dominant white is a widespread coat 62 color among domestic pig breeds, such as Landrace and Large White [1]. The dominant 63 white phen...

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“…Isolation and culture of peritoneal cell derived mast cells was performed as previously described (Meurer et al, 2016) with minor modifications. Briefly, 5 ml of PBS and 2 ml of air was injected into the peritoneal cavity of 14 week old mice.…”

Section: Isolation and Culture Of Peritoneal Cell Derived Mast Cellsmentioning

confidence: 99%

Functional Analysis of KIT Gene Structural Mutations Causing the Porcine Dominant White Phenotype Using Genome Edited Mouse Models

Sun

Qin

et al. 2020

Front. Genet.

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The dominant white phenotype in pigs is thought to be mainly due to a structural mutation in the KIT gene, a splice mutation (G > A) at the first base in intron 17 which leads to the deletion of exon 17 in the mature KIT mRNA. However, this hypothesis has not yet been validated by functional studies. Here, we created two mouse models, KIT D17/+ to mimic the splice mutation, and KIT Dup/+ to partially mimic the duplication mutation of KIT gene in dominant white pigs using CRISPR/Cas9 technology. We found that the splice mutation homozygote is lethal and the heterozygous mice have a piebald coat. Slightly increased expression of KIT in KIT Dup/+ mice did not confer the patched phenotype and had no obvious impact on coat color. Interestingly, the combination of these two mutations reduced the phosphorylation of PI3K and MAPK pathway associated proteins, which may be related to the impaired migration of melanoblasts observed during embryonic development that eventually leads to the dominant white phenotype.

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Isolation of Mature (Peritoneum-Derived) Mast Cells and Immature (Bone Marrow-Derived) Mast Cell Precursors from Mice

Cited by 56 publications

References 21 publications

Inhibitory effect of a new orally active cedrol-loaded nanostructured lipid carrier on compound 48/80-induced mast cell degranulation and anaphylactic shock in mice

Inhibitory effect of a new orally active cedrol-loaded nanostructured lipid carrier on compound 48/80-induced mast cell degranulation and anaphylactic shock in mice

Functional analysis of KIT gene structural mutations causing porcine dominant white phenotype by using genome edited mouse models

Functional Analysis of KIT Gene Structural Mutations Causing the Porcine Dominant White Phenotype Using Genome Edited Mouse Models

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