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Confluence-Associated Proliferation and Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cell (BMMSCs)

Author Affiliations

  • 1Department of Parasitology and Animal Diseases, Veterinary Research Division, National Research Center, Cairo, Egypt
  • 2Department of Parasitology and Animal Diseases, Veterinary Research Division, National Research Center, Cairo, Egypt
  • 3Department of Physiology, College of Veterinary Medicine, Cairo University, Giza, Egypt

Int. Res. J. Biological Sci., Volume 5, Issue (5), Pages 45-56, May,10 (2016)

Abstract

In cellular therapy field, the impact of confluence degree to harvest or differentiate BMMSCs and the effect created by cell-to-cell contact remains controversial. Therefore, the impact of 20, 50, 70, 80 and 100% confluences on BMMSCs proliferation properties, ERK and p-ERK proteins expression and glucose consumption rate was studied. Confluence-associated osteogenic differentiation efficiency was identified by determining calcium deposition, alizarin red staining, Alkaline phosphatase activity and osteopontin and osteocalcin genes expression. There was a correlation between confluence% and density. Viability was declined at the lower and higher confluences. The highest CFU-F, Brd-U uptake and population doubling were obtained at 80% confluence. ERK band intensity in 100% confluent BMMSCs was lower. Bands of p-ERK were highly detectable at 70% and 80% confluences. Glucose consumption rate at 70% and 80% confluences were higher than at 20% and 100% confluences. Although higher osteogenic differentiation appeared at 80% confluence, it was also extended at 100% confluence. Osteopontin gene expressed among all confluences while osteocalcin gene was highly expressed in 70% confluence. We concluded that the optimum seeding density for maximal expansion and harvesting purposes is 80% confluence and up to 100% confluence for osteogenic differentiation to trigger the process to be more cost effective.

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