TY - JOUR

T1 - Isolation, culturing and gene expression profiling of inner mass cells from stable and vulnerable carotid atherosclerotic plaques

AU - Novikova, Olga A.

AU - Nazarkina, Zhanna K.

AU - Cherepanova, Anna V.

AU - Laktionov, Petr P.

AU - Chelobanov, Boris P.

AU - Murashov, Ivan S.

AU - Deev, Roman V.

AU - Pokushalov, Evgeny A.

AU - Karpenko, Andrey A.

AU - Laktionov, Pavel P.

PY - 2018/8/1

Y1 - 2018/8/1

N2 - The connective tissue components that form the atherosclerotic plaque body are produced by the plaque inner mass cells (PIMC), located inside the plaque. We report an approach to isolate and culture cells from the connective tissue of stable and vulnerable human atherosclerotic plaques based on elimination of non-connective tissue cells such as blood and non-plaque intima cells with a lysis buffer. The resulting plaque cells were characterized by growth capacity, morphology, transcriptome profiling and specific protein expression. Plaque cells slowly proliferated for up to three passages unaffected by the use of proliferation stimulants or changes of culture media composition. Stable plaques yielded more cells than vulnerable ones. Plaque cell cultures also contained several morphological cellular types. RNA-seq profiles of plaque cells were different from any of the cell types known to be involved in atherogenesis. The expression of the following proteins was observed in cultured plaque cells: smooth muscle cells marker α-SMA, macrophage marker CD14, extracellular matrix proteins aggrecan, fibronectin, neovascularisation markers VEGF-A, CD105, cellular adhesion receptor CD31 and progenitor/dedifferentiation receptor CD34. Differential expression of several notable transcripts in cells from stable and vulnerable plaques suggests the value of plaque cell culture studies for the search of plaque vulnerability markers.

AB - The connective tissue components that form the atherosclerotic plaque body are produced by the plaque inner mass cells (PIMC), located inside the plaque. We report an approach to isolate and culture cells from the connective tissue of stable and vulnerable human atherosclerotic plaques based on elimination of non-connective tissue cells such as blood and non-plaque intima cells with a lysis buffer. The resulting plaque cells were characterized by growth capacity, morphology, transcriptome profiling and specific protein expression. Plaque cells slowly proliferated for up to three passages unaffected by the use of proliferation stimulants or changes of culture media composition. Stable plaques yielded more cells than vulnerable ones. Plaque cell cultures also contained several morphological cellular types. RNA-seq profiles of plaque cells were different from any of the cell types known to be involved in atherogenesis. The expression of the following proteins was observed in cultured plaque cells: smooth muscle cells marker α-SMA, macrophage marker CD14, extracellular matrix proteins aggrecan, fibronectin, neovascularisation markers VEGF-A, CD105, cellular adhesion receptor CD31 and progenitor/dedifferentiation receptor CD34. Differential expression of several notable transcripts in cells from stable and vulnerable plaques suggests the value of plaque cell culture studies for the search of plaque vulnerability markers.

KW - SMOOTH-MUSCLE-CELLS

KW - IDENTIFICATION

KW - PROGRESSION

KW - TRANSDIFFERENTIATION

KW - INFLAMMATION

KW - FIBRONECTIN

KW - APOPTOSIS

KW - BIOLOGY

KW - PLASMA

KW - Actins/metabolism

KW - Antigens, CD/metabolism

KW - Humans

KW - Macrophages/metabolism

KW - Atherosclerosis/genetics

KW - Male

KW - Plaque, Atherosclerotic/genetics

KW - Cell Proliferation/genetics

KW - Female

KW - Gene Expression Profiling/methods

KW - Carotid Arteries/metabolism

KW - Biomarkers/metabolism

KW - Transcriptome/genetics

KW - Vascular Endothelial Growth Factor A/metabolism

KW - Myocytes, Smooth Muscle/metabolism

KW - Aged

UR - http://www.scopus.com/inward/record.url?scp=85068835459&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0218892

DO - 10.1371/journal.pone.0218892

M3 - Article

C2 - 31242269

AN - SCOPUS:85068835459

VL - 14

SP - e0218892

JO - PLoS ONE

JF - PLoS ONE

SN - 1932-6203

IS - 6

M1 - e0218892

ER -