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Biology
成年小鼠泪液和下颌腺肌上皮细胞的分离
成年小鼠泪液和下颌腺肌上皮细胞的分离
JoVE Journal
Biology
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JoVE Journal Biology
Isolation of Myoepithelial Cells from Adult Murine Lacrimal and Submandibular Glands

成年小鼠泪液和下颌腺肌上皮细胞的分离

Full Text
8,598 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

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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.

泪腺 (LG) 有两种细胞类型表达α平滑肌肌动蛋白 (αSMA): 肌上皮细胞 (Mec) 和周周细胞。Mec 是外胚层的起源, 发现在许多腺体组织, 而周围的血管平滑肌细胞的内皮起源。该协议将 Mec 和 pericytes 与小鼠 Lg 隔离开来。

该协议是重要的,因为它允许分离和隔离两个光滑的肌肉细胞系,肌皮细胞和心状体。此方法通过细胞表面标记结合平滑肌肉细胞的遗传标记,以净化肌皮细胞和心状细胞的种群。该协议允许比较健康和患病的脑皮细胞的功能和再生能力,并处理这些细胞的基因表达研究。

乳腺细胞存在于其他外分泌腺体中,如乳腺、唾液和胰腺,这使得通过分离任何其他组织的脑皮细胞和周利特来适应此协议。要标记α平滑肌肉行为素或SMA表达细胞,注射三至四周大的塔莫西芬-可吸收阿尔法SMA驱动记者小鼠与100微升的塔莫西芬每20克体重内,每天两天一次。对于乳腺收集,在上次注射后两到三天,使用钳子轻轻拉一个腺体,同时用一把小剪刀的尖尖抓划腺周围的结缔组织,以释放腺体。

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