2010•Journal of Practical OrthopaedicsRequires access

Osteogenic Differentiation of Adipose-Derived Stem Cells from Rabbits in Vitro

Sun Xiao-tang

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Abstract

Objective To explore the differentiation of adipose-derived stem cells(ADSCs)from rabbits into osteogenic cells in vitro.Methods In this study,ADSCs were gained by collagenaseⅠdigestion of subcutaneous fat from the nape of New-Zealand white rabbit.Cell surface antigen CD44、CD45 were identified by immunofluorescence and cell auxodrome were drawn.The ADSCs were performed osteogenic and adipogenic differentiation respectively,following with texts related morphology,alkaline phosphatase assay,VonKossa and rathonum red O stain.ADSCs were divided into osteogenic induced-group and osteogenic noninduced-group.Alkaline phosphatase assay and calcium concentration of these two groups were checked and statistics from the check were analyzed at 1,2,3,4w after these two groups were both induced.Results This study got following results:a)ADSCs CD44 expression were positive and ADSCs CD45 expression negative;cell auxodromes were S shaped.b)Alkaline phosphatase assay,VonKossa and rathonum red O stain of ADSCs were all positive under the influence of osteogenic and adipogenic differentiation mediums.c)Alkaline phosphatase assays(ALP)were different of these two groups-ALP of induced-groups was higher than that of non-induced group at each different time(P0.05,n=5)and ALP of the induced-group was distinct in different tendency under different induction(P0.05,n=5).Calcium concentration were different of these two groups-calcium concentration of induced-groups was higher than that of non-induced group at each different time(P0.05,n=5)and calcium concentration of the induced group was distinct in different tendency under different induction(P0.05,n=5).Conclusion This study suggests that ADSCs can be isolated and cultured easily and have the property of fast generation and favourable osteogenic differentiation,which makes ADSCs ideal seed cells of bone tissue engineering.

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Objective To explore the differentiation of adipose-derived stem cells(ADSCs)from rabbits into osteogenic cells in vitro.Methods In this study,ADSCs were gained by collagenaseⅠdigestion of subcutaneous fat from the nape of New-Zealand white rabbit.Cell surface antigen CD44、CD45 were identified by immunofluorescence and cell auxodrome were drawn.The ADSCs were performed osteogenic and adipogenic differentiation respectively,following with texts related morphology,alkaline phosphatase assay,VonKossa and rathonum red O stain.ADSCs were divided into osteogenic induced-group and osteogenic noninduced-group.Alkaline phosphatase assay and calcium concentration of these two groups were checked and statistics from the check were analyzed at 1,2,3,4w after these two groups were both induced.Results This study got following results:a)ADSCs CD44 expression were positive and ADSCs CD45 expression negative;cell auxodromes were S shaped.b)Alkaline phosphatase assay,VonKossa and rathonum red O stain of ADSCs were all positive under the influence of osteogenic and adipogenic differentiation mediums.c)Alkaline phosphatase assays(ALP)were different of these two groups-ALP of induced-groups was higher than that of non-induced group at each different time(P0.05,n=5)and ALP of the induced-group was distinct in different tendency under different induction(P0.05,n=5).Calcium concentration were different of these two groups-calcium concentration of induced-groups was higher than that of non-induced group at each different time(P0.05,n=5)and calcium concentration of the induced group was distinct in different tendency under different induction(P0.05,n=5).Conclusion This study suggests that ADSCs can be isolated and cultured easily and have the property of fast generation and favourable osteogenic differentiation,which makes ADSCs ideal seed cells of bone tissue engineering.

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Available abstract

Objective To explore the differentiation of adipose-derived stem cells(ADSCs)from rabbits into osteogenic cells in vitro.Methods In this study,ADSCs were gained by collagenaseⅠdigestion of subcutaneous fat from the nape of New-Zealand white rabbit.Cell surface antigen CD44、CD45 were identified by immunofluorescence and cell auxodrome were drawn.The ADSCs were performed osteogenic and adipogenic differentiation respectively,following with texts related morphology,alkaline phosphatase assay,VonKossa and rathonum red O stain.ADSCs were divided into osteogenic induced-group and osteogenic noninduced-group.Alkaline phosphatase assay and calcium concentration of these two groups were checked and statistics from the check were analyzed at 1,2,3,4w after these two groups were both induced.Results This study got following results:a)ADSCs CD44 expression were positive and ADSCs CD45 expression negative;cell auxodromes were S shaped.b)Alkaline phosphatase assay,VonKossa and rathonum red O stain of ADSCs were all positive under the influence of osteogenic and adipogenic differentiation mediums.c)Alkaline phosphatase assays(ALP)were different of these two groups-ALP of induced-groups was higher than that of non-induced group at each different time(P0.05,n=5)and ALP of the induced-group was distinct in different tendency under different induction(P0.05,n=5).Calcium concentration were different of these two groups-calcium concentration of induced-groups was higher than that of non-induced group at each different time(P0.05,n=5)and calcium concentration of the induced group was distinct in different tendency under different induction(P0.05,n=5).Conclusion This study suggests that ADSCs can be isolated and cultured easily and have the property of fast generation and favourable osteogenic differentiation,which makes ADSCs ideal seed cells of bone tissue engineering.

Key concepts: Alkaline phosphatase, Adipose tissue, Adipogenesis, Collagenase, Calcium, Stem cell, CD44, Andrology

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