Isolation of Myoepithelial Cells from Adult Murine Lacrimal and Submandibular Glands

Tatiana Zyrianova; Liana  V. Basova; Helen Makarenkova

doi:10.3791/59602

JoVE Journal
Biology

A subscription to JoVE is required to view this content. Sign in or start your free trial.

JoVE Journal Biology

Isolation of Myoepithelial Cells from Adult Murine Lacrimal and Submandibular Glands

成人ネズミ涙腺および顎下腺からの筋上皮細胞の単離

Full Text

8,602 Views

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

Please note that some of the translations on this page are AI generated. Click here for the English version.

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.

涙腺 (LG) は、α平滑筋アクチン (αSMA): 筋上皮細胞 (MECs) および周皮細胞を発現する2つの細胞型を有する。MECs は外胚葉の起源であり、多くの腺組織に見られる一方、周皮細胞は endodermal 起源の血管平滑筋細胞である。このプロトコルは、マウス LGs から MECs および周皮細胞を分離する。

このプロトコルは、2つの平滑筋細胞株、筋上皮細胞および心細胞の分離および単離を可能にするので重要である。この方法は、細胞表面マーカーを介して平滑筋細胞の遺伝的標識を組み合わせて、筋上皮細胞および心細胞の集団を精製する。このプロトコルは、健康で病気の筋上皮細胞の機能と再生能力を比較し、遺伝子発現研究のためにこれらの細胞を処理することを可能にする。

筋上皮細胞は乳腺、唾液、膵臓などの他の外分泌腺に存在し、このプロトコルは他の組織からの筋上皮細胞および近縁細胞の単離によって適応することを可能にする。アルファ平滑筋アクチンまたはSMA発現細胞にラベルを付けるには、3〜4週齢のタモキシフェン誘導性アルファSMA駆動レポーターマウスを、1日1回腹腔内に10グラムの体重100マイクロリットルのタモキシフェンを2日間注入する。最後の注射の2〜3日後のラクリマル腺コレクションでは、ピンセットを使用して1つの腺を優しく引っ張ると同時に、小さなはさみの鋭い先端で腺の周りの結合組織を掻いて腺を解放します。

View the full transcript and gain access to thousands of scientific videos