Research output: Contribution to journal › Article › peer-review
Isolation, culturing and gene expression profiling of inner mass cells from stable and vulnerable carotid atherosclerotic plaques. / Novikova, Olga A.; Nazarkina, Zhanna K.; Cherepanova, Anna V. et al.
In: PLoS ONE, Vol. 14, No. 6, e0218892, 01.08.2018, p. e0218892.Research output: Contribution to journal › Article › peer-review
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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 -
ID: 20849356