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

Autor: Novikova OA; 'E. Meshalkin National Medical Research Center', Ministry of Health of the Russian Federation, Novosibirsk, Russia., Nazarkina ZK; Laboratory of Molecular Medicine, SB RAS Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia., Cherepanova AV; 'E. Meshalkin National Medical Research Center', Ministry of Health of the Russian Federation, Novosibirsk, Russia.; Laboratory of Molecular Medicine, SB RAS Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia., Laktionov PP; Laboratory of Genomics, SB RAS Institute of Molecular and Cellular Biology, Novosibirsk, Russia.; Novosibirsk State University, Novosibirsk, Russia., Chelobanov BP; Laboratory of Molecular Medicine, SB RAS Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia.; Novosibirsk State University, Novosibirsk, Russia., Murashov IS; 'E. Meshalkin National Medical Research Center', Ministry of Health of the Russian Federation, Novosibirsk, Russia., Deev RV; Ryazan State Medical University, Ryazan, Russia., Pokushalov EA; 'E. Meshalkin National Medical Research Center', Ministry of Health of the Russian Federation, Novosibirsk, Russia.; Laboratory of Molecular Medicine, SB RAS Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia., Karpenko AA; 'E. Meshalkin National Medical Research Center', Ministry of Health of the Russian Federation, Novosibirsk, Russia., Laktionov PP; 'E. Meshalkin National Medical Research Center', Ministry of Health of the Russian Federation, Novosibirsk, Russia.; Laboratory of Molecular Medicine, SB RAS Institute of Chemical Biology and Fundamental Medicine, Novosibirsk, Russia.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2019 Jun 26; Vol. 14 (6), pp. e0218892. Date of Electronic Publication: 2019 Jun 26 (Print Publication: 2019).
DOI: 10.1371/journal.pone.0218892
Abstrakt: 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.
Competing Interests: The authors have declared that no competing interests exist.
Databáze: MEDLINE
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