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Registration number: ПИ №ФС77-235В7,
8th March 2008. ISSN: 1815-445X
8th March 2008. ISSN: 1815-445X
Culturing of stromal cells derived from rat bone marrow in different origin I type collagen
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PDF1. The Saint Petersburg IP. Pavlov State Medical University, Saint Petersburg
2. The Saint Petersburg State University of Technology and Design, Saint Petersburg
3. The RAS Institute of Cytology, Saint Petersburg
2. The Saint Petersburg State University of Technology and Design, Saint Petersburg
3. The RAS Institute of Cytology, Saint Petersburg
Keywords:
bone marrow stromal cells, type I collagen, spreading, cytoskeleton, proliferation, differentiation
bone marrow stromal cells, type I collagen, spreading, cytoskeleton, proliferation, differentiation
Type I collagen is widely used in medicine and biotechnology owing to its ready
availability and good biological properties. The aim of this study was to
investigate proliferation (proliferation index), spreading (actin cytoskeleton,
spreading square] and osteogenic differentiation of the rat bone marrow stromal
cells (BMSCs] on the different molecular and fibrillar collagen substrates on
the surface and during three-dimensional cultivation in the collagen gels.
Collagen samples [1, 2, 3] were obtained from the skin of the sheep and young
bull by alkaline extraction. Control collagen [4] was obtained by acid
extraction from rat tail tendons. Fibrillar collagen on the surface was
prepared by subsequent threefold application of the molecular collagen layers
on the substrate. The capacity to form gels of the collagen samples in neutral
medium at 37?C was examined. BMSCs of the outbread rats were obtained by
centrifugation on the histopaque gradient with 1,077 g\ml density and were
cultured in the growth medium. Actin cytoskeleton of the spreaded cells was
stained with rhodamine-phalloidin. Distribution of the early osteogenic
differentiation marker — the alcaline phosphatase — was elicited
histochemically by staining of the cells with the standard solution containing
tetrazolium blue. In the course of the study was determined that not only
control collagen obtained from rat tails [4], but one collagen sample obtained
from the sheep skin [1] has the capacity to form gel. Hereafter we compared
cell actions on the fibrillar (gel-forming] [1 and 4] and molecular (not
gel-forming] [2 and 3] collagen samples. BMSCs had maximal spreading square,
proliferation index and alcaline phosphatase activity on the fibrillar
collagens [1 and 4]. Both collagens formed stable gels in which cells kept
osteogenic differentiation capacity until two weeks. Gel-forming sample of the
type I collagen obtained from the sheep skin can be recommended as a scaffold
for BMSCs transplantation.
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