Osteogenic differentiation of human umbilical cord mesenchymal stem cells induced with different concentrations of dexamethasone in vitro

doi:10.3969/j.issn.2095-4344.2013.23.004

Abstract

Abstract:

BACKGROUND: Under the combination effect of dexamethasone, vitamin C and β-glycerophosphate, human umbilical cord mesenchymal stem cells can differentiate into osteoblasts. Dexamethasone is a glucocorticoid hormone, and its concentration can affect the osteogenic differentiation of human umbilical cord mesenchymal stem cells.
OBJECTIVE: To explore the optimal concentrations of dexamethasone in the osteogenic differentiation of human umbilical cord mesenchymal stem cells.
METHODS: Human umbilical cord mesenchymal stem cells were isolated from the normal full-term newborn umbilical cord by explant method. The passage 3 human umbilical cord mesenchymal stem cells were used for routine cell culture and induced differentiation experiments. The expression of surface markers of stem cells was detected with flow cytomerey, and the human umbilical cord mesenchymal stem cells were identified by osteogenic differentiation and adipogenic differentiation. The changes of cell morphology were observed under the inverted phase contrast microscope in the process of osteogenic differentiation. The induced medium for osteogenic differentiation containing 1×^10-8 mol/L, 5×10^-8 mol/L and 1×10^-7 mol/L dexamethasone was used for the osteogenic induction of umbilical cord mesenchymal stem cells. Tetracycline hydrochloride fluorescence and Von Kossa staining were applied to observe the formation of calcium nodules. Reverse transcriptase-PCR was used to detect the expression of osteopontin gene.
RESULTS AND CONCLUSION: The form of human umbilical cord mesenchymal stem cells separated by explant method was uniform, spindle and growing in parallel and vortex-like shape. The flow cytometry results showed that CD29, CD73 and CD90 were positive, and CD31, CD34 and HLA-DR were negative. Intracellular lipid droplets could be observed after adipogenic differentiation, and the formation size and quantity of calcium nodules were enhanced as the concentration of dexamethasone increased. The expression level of osteopontin gene could be seen in three concentrations group, and the expressions were enhanced with the increasing of dexamethasone concentration from reverse transcriptase-PCR results. Human umbilical cord mesenchymal stem cells can be effectively induced to differentiate into osteobalsts when the medium contains 1×10^-7 mol/L dexamethasone.
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Key words: stem cells, umbilical cord/umbilical cord blood stem cells, dexamethasone, optimal concentration, osteogenic differentiation, adipogenic differentiation, calcium nodules, osteopontin, cell culture, cell identification, cell morphology, stem cell photographs-containing paper

CLC Number:

R394.2

Hong Jing-xin, Liu Jian, Li Lin-fang, Han Jun-ling. Osteogenic differentiation of human umbilical cord mesenchymal stem cells induced with different concentrations of dexamethasone in vitro[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(23): 4204-4211.

Figures/Tables 7

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