Isolation of Myoepithelial Cells from Adult Murine Lacrimal and Submandibular Glands

Tatiana Zyrianova; Liana  V. Basova; Helen Makarenkova

doi:10.3791/59602

Isolamento de células mioepiteliais das glândulas lacrimal e submandibular de Murina adulta

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Isolation of Myoepithelial Cells from Adult Murine Lacrimal and Submandibular Glands

Isolamento de células mioepiteliais das glândulas lacrimal e submandibular de Murina adulta

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07:15 min

June 11, 2019

DOI: 10.3791/59602-v

Tatiana Zyrianova¹, Liana V. Basova¹, Helen Makarenkova¹

¹Department of Molecular Medicine,The Scripps Research Institute

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Overview

This study presents a protocol for the isolation and separation of two smooth muscle cell types from the lacrimal gland: myoepithelial cells (MECs) and pericytes. Using genetic labeling, the method enables purification for comparative analysis of healthy and diseased cells, thereby contributing to our understanding of cell function and regenerative abilities.

Key Study Components

Research Area

Cell biology
Regenerative medicine
Exocrine gland research

Background

MECs and pericytes play crucial roles in glandular function and vascular stability.
This protocol is adaptable for isolating similar cell types from other exocrine glands.
Understanding their functions can provide insights into various biological processes and diseases.

Methods Used

Genetic labeling and purification of myoepithelial cells and pericytes.
Murine model with tamoxifen-inducible alpha SMA driven reporter mice.
Fluorescence-activated Cell Sorting (FACS) for cell analysis.

Main Results

Successfully isolated and labeled MECs and pericytes from murine lacrimal glands.
Facilitated downstream applications including cell culture and gene expression studies.
Demonstrated differences in cell morphology and labeling efficiency.

Conclusions

The study provides a valuable protocol for the isolation of specific cell types within the lacrimal gland.
This advancement may enhance future research related to exocrine glands and their cellular functions.

Frequently Asked Questions

What are the key benefits of this isolation protocol?

It allows the purification of specific cell types for detailed analysis of their functions and regenerative abilities.

Can this method be used for other tissues?

Yes, the protocol can be adapted for isolation from other exocrine glands.

What is the role of myoepithelial cells?

Myoepithelial cells assist in glandular secretion and contractile functions.

What are pericytes and their significance?

Pericytes are essential for vascular stability and regulation of blood flow in capillaries.

What applications can the isolated cells be used for?

The cells can be utilized for in vivo/in vitro experiments, including cell culture and molecular studies.

How is cell labeling achieved in this protocol?

Cell labeling is accomplished through injection of tamoxifen to activate the desired reporting mechanism.

What technologies were critical for this study?

Fluorescence microscopy and FACS were essential for cell analysis and sorting.

A glândula lacrimal (LG) tem dois tipos de células expressando α-actínio do músculo liso (αSMA): células mioepiteliais (MECs) e pericytes. Os MECS são de origem ectodérmica, encontrados em muitos tecidos glandulares, enquanto os pericitos são células musculares lisas vasculares de origem endodérmica. Este protocolo isola MECS e pericitos de LGS murino.

Este protocolo é significativo porque permite a separação e isolamento de duas linhas de células musculares lisas, as células mioepitheliais e pericítas. Este método combina rotulagem genética de células musculares lisas através de marcadores de superfície celular para purificar populações de células mioepiteliais e pericítos. Este protocolo permite comparar a função e as habilidades regenerativas das células mioepitheliais saudáveis e doentes e processar essas células para estudos de expressão genética.

As células mioepitheliais existem em outras glândulas exócrinas como mamária, salivar e pâncreas, o que permite que este protocolo seja adaptado pelo isolamento de células mioepiteliais e pericítos de qualquer outro tecido. Para rotular as células de expressão do músculo alfa liso ou SMA, injete de três a quatro semanas de idade, camundongos repórteres alfa sma conduzidos por tamoxifen com 100 microliters de tamoxifen por 20 gramas de peso corporal intraperitonel uma vez por dia durante dois dias. Para a coleta de glândula lacrimal, dois a três dias após a última injeção, use pinças para puxar suavemente uma glândula e, ao mesmo tempo, coçar o tecido conjuntivo ao redor da glândula com a ponta afiada de um par de tesouras pequenas para libertar a glândula.

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