Abstract:

BACKGROUND: Smooth muscle progenitor cells (SPCs) from human peripheral blood have the capacity to differentiate into smooth muscle cells.
OBJECTIVE: To investigate the method of in vitro purifying and culturing SPCs, and to observe differentiation and growth of SPCs.
METHODS: Mononuclear cells were isolated from adult human peripheral blood by density gradient centrifugation. After 12 days of culture in vitro, SPCs were identified and sorted by flow cytometry. SPCs were then induced to differentiate and amplify. Inverted microscope was used to observe morphological changes. Smooth muscle cells specific markers (α-SMA, Calponin, SM-MHC) were then checked with immunofluorescent staining and Western blotting. In addition, Cell growth curve of culture was drawn.
RESULTS AND CONCLUSION: Four days later, cell colonies were presented. Twelve days later, most adherent cells were showing spindle-shape. CD14/CD105 double positive cells were SPCs, which accounted for (71.8±7.2)% of adherent cells. Twenty-eight days later, cells arranged in order, showing whirlpool-shape. On day 28, immunostaining of adherent cells was positive for α-SMA. Expression of calponin and SM-MHC was found at 14 d and 21 d with Western blotting respectively, and they gradually enhanced. Growth curve of SPCs showed that increased logarithmic phase started on the 6th day, and platform phase started on the 12th days. Results indicated that SPCs could be harvested through culturing mononuclear cells from adult human peripheral blood. They could maintain the ability of proliferation, and differentiate into smooth muscle cells in vitro. So, SPCs could be selected as the seed cells.