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BACKGROUND:There are various methodstoinduceadipogenic differentiation ofbone marrow mesenchymal stem cels, and the main componentfor adipogenic induction isindomethacin or rosiglitazone. However, there is a lack of comparative studyonthe induction efficiency and mechanism among these methods. OBJECTIVE:Tocompare the adipogenic responses ofhuman bone marrow mesenchymal stem celsto different induction methods, and to analyze the mechanismunderlyingdifferent induction efficiency. METHODS:After isolation and purification,the adipogenic abilitiesof human bone marrow mesenchymal stem cels in threedifferentculture systemswere comparedby oil red O staining and lipogenic geneassay. At 0, 1, 3 and 7 days of adipogenensis, mRNA expressionsof PPARγ, C/EBPα, Adiponectin and Leptin were detected.At7 daysofadipogenensis, protein expressionsof PPARγ and C/EBPβ were detectedby western blot assay,andeffects ofDIMIversusDIMRonphosphorylationofPPARγatSer273were compared. RESULTS AND CONCLUSION:Findings from oil red O staining andreal-time PCRshowedthat DIMR significantlyinducedadipogenicdifferentiation of bonemarrow mesenchymal stem cels compared with DIM and DIMI at 7 daysofinduction. Western blot showed thattheprotein expressionsof PPARγ and C/EBPβ in the DIMIgroupwere significantly higher than those in the DIMRand DIM at 7days ofinduction. In addition, the ratio ofPPARγphosphorylation atSer273was lowerin the DIMR group thantheDIMI group.To conclude,DIMR has the most potential to induce early adipogenesis ofhumanbone marrow mesenchymal stem cels by weakening the phosphorylationof PPARγ-Ser273.
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Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. Mature purification process was established and recombinant protein SMB(PRG4), with high-level expression was purified. By using size-exclusion chromatogaraphy and dynamic light scattering, we found that the recombinant protein self-aggregate to dimeric form. Structure prediction and non-reducing electrophoresis revealed that SMB(PRG4), was a non-covalently bonded dimer.
Subject(s)
Glycosylation , Protein Multimerization , Proteoglycans , Chemistry , Recombinant Proteins , Chemistry , Somatomedins , ChemistryABSTRACT
BACKGROUND:An effective freezing-thawing technique is crucial for the clinical application of human umbilical cord mesenchymal stem cells(UC-MSCs).OBJECTIVE:To investigate biological characteristics of UC-MSCs after cryopreservation.METHODS:UC-MSCs were isolated from human umbilical cord and frozen in liquid nitrogen.The survival rate and the suppressive effect of γ-interferon(IFN-γ)of cryopreserved-thawed and fresh human UC-MSCs were compared.Furthermore,the multiple potentials and phenotype of UC-MSCs were estimated after cryopreservation.RESULTS AND CONCLUSION:There was no significant difference between cryopreserved-thawed and fresh human UC-MSCs on the survival rate and the suppressive effect of IFN-γ of peripheral blood mononuclear cells(PBMCs).After cryopreservation,human UC-MSCs had the potential differentiation and the phenotype of mesenchymal stem cells.
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BACKGROUND:It is not certain that whether human adipose-derived mesanchymal stem cells express telomerase reverse transcriptase (TERT).OBJECTIVE:To investigate whether human adipose-derived mesenchymal stem cells express telomerase reverse transcriptase or not.DESIGN,TIME AND SETTING:The cytology in vitro experiments were performed at TEDA Life and Technology Research Center,Institute of Hematology and Blood Diseases Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College in August 2009.MATERIALS:Adult adipose tissue was obtained from 5 healthy donors undergoing liposuction at the Tianjin Yili Medical Cosmetology Plastics Outpatient.Hela cells were supplied by the Cell Center of Basic Medical Sciences,Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences.METHODS:Human adipose-derived mesenchymal stem cells were isolated from 20 mL human edipose tissue of each healthy donor by collagenase digestion.Cells were digested by trypsin when 80% confluency.Hela cells served as controls,and treated with 1640 medium containing 10% fetal bovine serum,and then digested using trypsin when 80% confluency.MAIN OUTCOME MEASURES:Morphology,phenotype and differentiation of human adipose-derived mesenchymal stem cells.TERT of human adipose-derived mesenchymal stem calls were detected by reverse transcription-polymerase chain reaction (RT-PCR).Telomerase activity was measured by telomeric repeat amplification protocol assay ELISA (TRAP-ELISA).RESULTS:Human adipose-derived mesenchymal stem cells adhered to the flask,presented spindle shape,and whirlpool-shape when cell density was large.Human adipose-derived mesenchymal stem cells were labeled positively for CD73,CD90 and CD105,but negatively for CD19,CD34,CD11b,CD45,and HLA-DR.Following adipogenic induction,oil red O staining showed significant red oil drop.Following ostesgenic induction,Von Kossa staining showed black particles,which indicated calcium deposition.Human adipose-derived mesenchymal stem cells at P1,P4 and P7 did not express telomerase reverse transcriptase gane.Human adipose-derived mesenchymal stem cells from 5 different donors did not expression telomerase reverse transcriptase gane.Hela cells had telomerase activation,but human adipose-derived mesanchymal stem cells did not express telomerase activation.CONCLUSION:Human adipose-derived mesanchymal stem cells do not express mRNA of hTERT,nor the activity of telomerase reverse transcriptase.
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Objective To evaluate the value of X-ray imaging in the diagnosis of multiple myeloma.Methods The findings of X-ray plain films of 136 patients with multiple myeloma were retrospectively analysed.Results Abnormal radiological changes were found in 117 of 136 patients (86.03%),of which 84 cases(61.76%) were located in skull,61 cases(44.85%) in spinal column,47(34.56%)in pelvis,51(37.5%)in humerus,41(30.15%)in ribs,29 (21.32%)in femur.In addition,clavical was involved in 15 cases,scapula in 13,radius in 9,tibia in 9,fibula in 4 and ulna in 3.Conclusion X-ray plain film is of important effect on the diagnosis of multiple myeloma.
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Objective To detect the expression of Zbtb7 gene in Phorbol esters(PMA) induced differentiation of leukemia cell lines.Methods 50 nmol/L PMA was used to induce the differentiation of U937 and K562 cell lines.The expression of Zbtb7 was detected by real-time PCR in the cells after induction on day 0,1,3 and 5,respectively. Results The expression of Zbtb7 in leukemia cell lines was markedly enhanced after treated by PMA(P