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Isolation and culture of vascular smooth muscle cells from rat placenta
Author(s) -
He Chunyu,
Wang Meili,
Yan Zi,
Zhang Suli,
Liu Huirong
Publication year - 2019
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.27721
Subject(s) - calponin , vascular smooth muscle , placenta , microbiology and biotechnology , intracellular , angiotensin ii , chemistry , smooth muscle , isolation (microbiology) , biology , protein kinase c , biochemistry , actin , endocrinology , kinase , fetus , receptor , pregnancy , bioinformatics , genetics
We developed a new separation method for isolating placental vascular smooth muscle cells (PVSMCs) from a rat in this study. Our method used the magnetic force between a magnet and ferrous ferric oxide (Fe 3 O 4 ) to make the separation and extraction processes easier and more efficient. From the first to sixth generation, the cells isolated using this protocol were identified as smooth muscle cells (SMCs) by their immunoreactivity to the SMC markers and by the “hill and valley” morphology. PVSMCs were exposed to angiotensin II (1 μmol/L) and resulted in sharply increased intracellular Ca 2+ concentration. Furthermore, activation of protein kinase C (PKC) increased concomitantly with a decrease in calponin expression. These results indicate that the isolated cells had biological activity. Our method of isolating PVSMCs from rat leads to isolation of cultured cells with activity and high purity. The approach will be useful in research studies on placental vascular diseases.