Isolation and Purification of Murine Cardiac Pericytes
Author(s) -
Linda L. Lee,
Aarif Y. Khakoo,
Vishnu Chintalgattu
Publication year - 2019
Publication title -
journal of visualized experiments
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.596
H-Index - 91
ISSN - 1940-087X
DOI - 10.3791/59571
Subject(s) - cd146 , cell sorting , microbiology and biotechnology , angiogenesis , cd31 , pericyte , biology , flow cytometry , primary cell , cd34 , cell , chemistry , endothelial stem cell , stem cell , cancer research , biochemistry , in vitro
Pericytes, perivascular cells of microvessels and capillaries, are known to play a part in angiogenesis, vessel stabilization, and endothelial barrier integrity. However, their tissue-specific functions in the heart are not well understood. Moreover, there is currently no protocol utilizing readily accessible materials to isolate and purify pericytes of cardiac origin. Our protocol focuses on using the widely used mammalian model, the mouse, as our source of cells. Using the enzymatic digestion and mechanical dissociation of heart tissue, we obtained a crude cell mixture that was further purified by fluorescence activating cell sorting (FACS) by a plethora of markers. Because there is no single unequivocal marker for pericytes, we gated for cells that were CD31 - CD34 - CD45 - CD140b + NG2 + CD146 + . Following purification, these primary cells were cultured and passaged multiple times without any changes in morphology and marker expression. With the ability to regularly obtain primary murine cardiac pericytes using our protocol, we hope to further understand the role of pericytes in cardiovascular physiology and their therapeutic potential.
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