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Updates in processing of anterior fat pad aspirate for amyloid (with video and sketches).

ABSTRACT: Fat pad aspiration is a commonly used method for detecting amyloid in tissue. Amyloid is detected in the small blood vessels of the aspirated adipose tissue. Optimum evaluation of amyloid with electron microscopy requires at least 15 blood vessels in the fat pad aspirate. The presence of a significant proportion of adipocytes in the aspirate dilutes the fibrovascular portion. This may compromise the evaluation for amyloid with electron microscopy and in FFPE with proteomic studies by mass spectroscopy for confirmation of the amyloid subtype. This video article describes the updated protocol for processing the anterior fat pad aspirate. It demonstrates how to remove the interference of blood and fatty component in the fat pad aspirate performed by the previously reported procedure.

SUBMITTER: Shidham VB

PROVIDER: S-EPMC7568089 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Updates in processing of anterior fat pad aspirate for amyloid (with video and sketches).

Shidham Vinod B VB

CytoJournal 20200620

Fat pad aspiration is a commonly used method for detecting amyloid in tissue. Amyloid is detected in the small blood vessels of the aspirated adipose tissue. Optimum evaluation of amyloid with electron microscopy requires at least 15 blood vessels in the fat pad aspirate. The presence of a significant proportion of adipocytes in the aspirate dilutes the fibrovascular portion. This may compromise the evaluation for amyloid with electron microscopy and in FFPE with proteomic studies by mass spectr ...[more]

PMID: 33093851

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Project description:BackgroundTo investigate the in vitro and in vivo anti-inflammatory/anti-fibrotic capacity of IFP-MSC manufactured as 3D spheroids. Our hypothesis is that IFP-MSC do not require prior cell priming to acquire a robust immunomodulatory phenotype in vitro in order to efficiently reverse synovitis and IFP fibrosis, and secondarily delay articular cartilage damage in vivo.MethodsHuman IFP-MSC immunophenotype, tripotentiality, and transcriptional profiles were assessed in 3D settings. Multiplex secretomes were assessed in IFP-MSC spheroids [Crude (non-immunoselected), CD146+ or CD146- immunoselected cells] and compared with 2D cultures with and without prior inflammatory/fibrotic cell priming. Functionally, IFP-MSC spheroids were assessed for their immunopotency on human PBMC proliferation and their effect on stimulated synoviocytes with inflammation and fibrotic cues. The anti-inflammatory and anti-fibrotic spheroid properties were further evaluated in vivo in a rat model of acute synovitis/fat pad fibrosis.ResultsSpheroids enhanced IFP-MSC phenotypic, transcriptional, and secretory immunomodulatory profiles compared to 2D cultures. Further, CD146+ IFP-MSC spheroids showed enhanced secretory and transcriptional profiles; however, these attributes were not reflected in a superior capacity to suppress activated PBMC. This suggests that 3D culturing settings are sufficient to induce an enhanced immunomodulatory phenotype in both Crude and CD146-immunoselected IFP-MSC. Crude IFP-MSC spheroids modulated the molecular response of synoviocytes previously exposed to inflammatory cues. Therapeutically, IFP-MSC spheroids retained substance P degradation potential in vivo, while effectively inducing resolution of inflammation/fibrosis of the synovium and fat pad. Furthermore, their presence resulted in arrest of articular cartilage degradation in a rat model of progressive synovitis and fat pad fibrosis.Conclusions3D spheroids confer IFP-MSC a reproducible and enhanced immunomodulatory effect in vitro and in vivo, circumventing the requirement of non-compliant cell priming or selection before administration and thereby streamlining cell products manufacturing protocols.

Dataset Information

Updates in processing of anterior fat pad aspirate for amyloid (with video and sketches).

Publications

Updates in processing of anterior fat pad aspirate for amyloid (with video and sketches).

Similar Datasets

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