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( Naveen Kumar Mekala ),( Rama Raju Baadhe ),( Sreenivasa Rao Parcha ),( Prameela Devi Y ) 한국조직공학·재생의학회 2012 조직공학과 재생의학 Vol.9 No.5
Mononuclear cells isolated from human umbilical cord blood were differentiated in to spindle shaped fibrous mesenchymal stem cells (MSCs) in DMEM with 10% fetal bovine serum. These progenitor cells were further examined for their ability to differentiate into osteoblasts by culturing them in osteogenic differentiation media (Stem Pro(R)). Calcium mineralization assay, alizarin red S, von kossa staining confirmed that MSCs from cord blood were capable of mineralization when they were cultured in osteogenic medium. The adhesion studies shown that MSCs have greater affinity towards fibronectin (FN) coated dishes, compared to albumin (Alb) coated dishes due to CD29and CD44 surface receptors.
Naveen Kumar Mekala,Rama Raju Baadhe,Sreenivasa Rao Parcha 한국조직공학과 재생의학회 2013 조직공학과 재생의학 Vol.10 No.4
Mesenchymal stem cells (MSCs) isolated from umbilical card blood were capable of differentiated in to multiple mesenchymal cell lineages. The objective of the current study was to establish a reproducible system for the in vitro osteogenic differentiation of human MSCs, and to characterize the effect of changes in the microenvironment upon differentiation. In our previous studies, MSCs cultured in varying concentrations of L-ascorbic acid (50µM to 500µM) had shown the various levels of osteoblast differentiation, as determined by morphology, alkaline phosphatase activity, modulations in osteocalcin mRNA expression, and mineralized extracellular matrix (ECM) deposition holding inorganic hydroxyapatite (HA) and Tricalcium phosphate (TCP). In present study, we noticed a study increase in APase activity up to 250µM L-ascorbic acid and by end of 16 days, cells gained cuboidal morphology, which is characteristic feature of osteoblasts. The intense sharp difractograms (peaks) between 30o-35o demonstrate the deposition of HA and TCP, which are major inorganic components of bone. The reproducible results of our studies provided a useful model for evaluating the factors responsible for the stepwise progression of cells from undifferentiated precursors to secretary osteoblasts, and eventually terminally differentiated osteocytes.