Expansion and Adipogenesis Induction of Adipocyte Progenitors from Perivascular Adipose Tissue Isolated by Magnetic Activated Cell Sorting

Kyan Thelen; Nadia Ayala-Lopez; Stephanie W. Watts; G. Andres Contreras

doi:10.3791/55818

Expansão e adipogênese Indução de progenitores de adipócitos do tecido adiposo perivascular isola...

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Expansion and Adipogenesis Induction of Adipocyte Progenitors from Perivascular Adipose Tissue Isolated by Magnetic Activated Cell Sorting

Expansão e adipogênese Indução de progenitores de adipócitos do tecido adiposo perivascular isolado pela classificação de células ativadas magnéticas

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08:28 min

June 30, 2017

DOI: 10.3791/55818-v

Kyan Thelen¹, Nadia Ayala-Lopez², Stephanie W. Watts², G. Andres Contreras¹

¹Department of Large Animal Clinical Sciences,Michigan State University, ²Department of Pharmacology and Toxicology,Michigan State University

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Overview

This study presents a method for isolating Adipocyte Progenitor Cells (APCs) from Perivascular Adipose Tissue (PVAT) using Magnetic-activated Cell Sorting (MCS). This technique enhances the yield of APCs per gram of adipose tissue compared to traditional Fluorescence-Activated Cell Sorting (FACS).

Key Study Components

Area of Science

Cell Biology
Adipose Tissue Research
Stem Cell Biology

Background

Adipocyte progenitor cells are crucial for understanding adipose tissue function.
Perivascular adipose tissue plays a significant role in metabolic processes.
Isolation techniques are essential for studying specific cell populations.
Magnetic-activated Cell Sorting offers advantages over traditional methods.

Purpose of Study

To develop a reliable method for isolating APCs from PVAT.
To facilitate the differentiation of APCs into mature adipocytes.
To analyze the viability and potential of isolated APCs.

Methods Used

Magnetic-activated Cell Sorting (MCS) for cell isolation.
Preparation of Krebs-Ringer bicarbonate buffer (KRBB) for cell handling.
Incision and tissue collection from rat models.
Assessment of cell morphology and viability post-isolation.

Main Results

Increased yield of APCs per gram of adipose tissue using MCS.
Minimal impact on cell morphology and viability during isolation.
Successful differentiation of isolated APCs into mature adipocytes.
Enhanced understanding of specific APC populations in PVAT.

Conclusions

The MCS method is effective for isolating APCs from PVAT.
This technique can advance research on adipocyte biology.
Future studies can leverage these findings for metabolic research.

Frequently Asked Questions

What is the significance of isolating APCs?

Isolating APCs allows researchers to study their role in adipose tissue and metabolic health.

How does MCS compare to FACS?

MCS provides a higher yield of APCs with less impact on cell viability and morphology.

What are the applications of this research?

This research can inform studies on obesity, diabetes, and metabolic disorders.

What are the next steps after isolating APCs?

The next steps include inducing differentiation into mature adipocytes and studying their function.

Can this method be applied to other tissues?

While this method is optimized for PVAT, similar techniques may be adapted for other tissues.

What precautions should be taken during the procedure?

Ensure proper anesthesia and surgical techniques to minimize stress and injury to the animal.

Aqui, relatamos um método para o isolamento das populações de células progênitentes de adipócitos (APC) do tecido de tecido adiposo perivascular (PVAT) usando a classificação celular celular ativada por magnética (MCS). Este método permite um maior isolamento de APC por grama de tecido adiposo quando comparado à Classificação de células ativadas por fluorescência (FACS).

Os objetivos gerais desses protocolos de isolamento e diferenciação são obter células progenitoras de adipócitos a partir de tecidos adiposos perivasculares e induzir sua diferenciação em adipócitos maduros. Este método permite a análise de populações de células progenitoras de adipócitos específicas e definidas do tecido adiposo perivascular, com impacto mínimo em sua morfologia, viabilidade e potencial de proliferação e diferenciação. A principal vantagem dessa técnica é que ela permite um maior isolamento de células apresentadoras de antígenos por grama de tecido adiposo, em comparação com a classificação de células ativadas por fluorescência.

Depois de confirmar a falta de resposta ao beliscão do dedo do pé, faça uma incisão vertical na linha média ao longo do esterno até a área perineal do rato, para acessar a cavidade abdominal. Exponha a artéria mesentérica superior, os pequenos vasos de resistência mesentérica e a aorta torácica e corte todas as conexões com o mesentério e a aorta. Colete o tecido adiposo gonadal e transfira as bolsas de gordura isoladas para um novo recipiente de tampão de bicarbonato de Krebs-Ringer, ou solução de KRBB, suplementado com montes de 10 milimolares.

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