RESUMO
To investigate the effect of hSCGF-alpha on human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs), we obtained hSCGF-alpha using genetic engineering, hSCGF-alpha gene was amplified from hUCMSCs cDNA using two-step PCR and was inserted into pET-28a(+) plasmid vector. Induced by IPTG at 20 degrees Celsius for 24 h, the fusion protein expressed in E. coli BL21 (DE3) was mainly existing in soluble form. The recombinant hSCGF-a was purified using NI-NTA affinity chromatography and the purity was up to 90%. The colony forming test revealed that combined use hSCGF-alpha and rmGM-CSF (recombinant murine GM-colony stimulating factor, rmGM-CSF) had granulocyte/macrophage (GM) promoting effects on murine bone marrow GM progenitor. In addition, the results indicated that hSCGF-alpha and rhGM-CSF had stimulatory effect on hUCMSCs and their synergetic effect was the strongest.
Assuntos
Humanos , Proliferação de Células , Células Cultivadas , Clonagem Molecular , Sinergismo Farmacológico , Escherichia coli , Genética , Metabolismo , Fator Estimulador de Colônias de Granulócitos e Macrófagos , Farmacologia , Células-Tronco Mesenquimais , Biologia Celular , Metabolismo , Proteínas Recombinantes de Fusão , Genética , Farmacologia , Fator de Células-Tronco , Genética , Cordão Umbilical , Biologia CelularRESUMO
BACKGROUND:Culture condition,isolation method and efficiency are different in reported human umbilical cord-derived mesenchymal stem cells,which lack of unified identification standards.Therefore,it is necessary to establish a high-efficiency and economical culture system for human umbilical cord-derived mesenchymal stem calls(hUCMSCs).OBJECTIVE:To isolate hUCMSCs and induced differentiate into adipocytes and osteblasts.METHODS:The hUCMSCs were isolated form human umbilical cord by tissue adherence and digested with collagenase.The morphology,proliferation and immunophenotype of the 3rd passage cells were analyzed,and then cells were induced to osteogenic and adipogenic differentiation in vitro.RESULTS AND CONCLUSION:The hUCMSCs isolated from human umbilical cord by tissue adherence and digested with collagenase could be cultured and proliferated in vitro.Flow cytometry analysis revealed that the hUCMSCs were positive for CD29 CD44,CD59,CD105,but were negative for CD40,CD86 and HLA-DR.These calls could be induced to differentiate into adipocytes and osteblasts under proper inducing conditions.The hUCMSCs retained the appearance and phenotype even after being expanded more than 40 passages in vitro.This confirmed that the existence of MSCs in human umbilical cord and they had the capacity of differentiating into adipocytes and osteblasts.
RESUMO
OBJECTIVE:To prepare core-shell nanocapsules loading nicardipine hydrochloride and to investigate its pharmaceutical characteristics. METHODS: Core-shell nanocapsules were prepared using layer-by-layer electrostatic self-assembly technique. The indexes including the shape and particle size and the loaded drug amount of the nanocapsules were evaluated, and its accumulative release rates in artificial gastric juice and intestinal juice were computed and compared with those of its crude drug. RESULTS: The results showed that the nanocapsules were spherical with a mean particle size of 200 nm and a maximum loaded drug amount of 2.512%. The drug release rate within 12 h reached 18.64% in artificial gastric juice and 70% in artificial intestinal juice, whereas within 3 h the drug release rate of its crude drug in artificial intestinal juice reached 87%. CONCLUSIONS: The prepared core-shell nanocapsules containing nicardipine hydrochloride had a good pharmaceutical property.