从成年小鼠泪腺和颌下腺中分离肌上皮细胞。
Isolation of Myoepithelial Cells from Adult Murine Lacrimal and Submandibular Glands.
作者信息
Zyrianova Tatiana, Basova Liana V, Makarenkova Helen
机构信息
Department of Molecular Medicine, The Scripps Research Institute.
Department of Molecular Medicine, The Scripps Research Institute;
出版信息
J Vis Exp. 2019 Jun 11(148). doi: 10.3791/59602.
The lacrimal gland (LG) is an exocrine tubuloacinar gland that secretes an aqueous layer of tear film. The LG epithelial tree is comprised of acinar, ductal epithelial, and myoepithelial cells (MECs). MECs express alpha smooth muscle actin (αSMA) and have a contractile function. They are found in multiple glandular organs and are of ectodermal origin. In addition, the LG contains SMA+ vascular smooth muscle cells of endodermal origin called pericytes: contractile cells that envelop the surface of vascular tubes. A new protocol allows us to isolate both MECs and pericytes from adult murine LGs and submandibular glands (SMGs). The protocol is based on the genetic labeling of MECs and pericytes using the SMA:Rosa26-TdTomato mouse strain, followed by preparation of the LG single-cell suspension for fluorescence activated cell sorting (FACS). The protocol allows for the separation of these two cell populations of different origins based on the expression of the epithelial cell adhesion molecule (EpCAM) by MECs, whereas pericytes do not express EpCAM. Isolated cells could be used for cell cultivation or gene expression analysis.
泪腺（LG）是一种外分泌性管泡状腺，分泌泪膜的水液层。泪腺上皮树由腺泡、导管上皮和肌上皮细胞（MECs）组成。MECs表达α平滑肌肌动蛋白（αSMA）并具有收缩功能。它们存在于多个腺器官中，起源于外胚层。此外，泪腺含有起源于内胚层的SMA+血管平滑肌细胞，称为周细胞：包裹血管表面的收缩细胞。一种新方法使我们能够从成年小鼠的泪腺和下颌下腺（SMG）中分离出MECs和周细胞。该方法基于使用SMA:Rosa26-TdTomato小鼠品系对MECs和周细胞进行基因标记，然后制备泪腺单细胞悬液用于荧光激活细胞分选（FACS）。该方法允许根据MECs对上皮细胞粘附分子（EpCAM）的表达来分离这两种不同起源的细胞群体，而周细胞不表达EpCAM。分离出的细胞可用于细胞培养或基因表达分析。
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