Heterogeneity of immunomodulatory function of exosomes derived from human umbilical cord mesenchymal stem cells / 军事医学

Objective To investigate the heterogeneity of immunomodulatory function of exosomes secreted from human umbilical cord-derived mesenchymal stem cells(hUC-MSC).Methods Five different hUC-MSCs lines were isolated and cultured.Exosomes were isolated from the supernatant.The expression of specific surface markers CD9,CD63,CD81 and CD44 was detected by flow cytometry.The protein concentration of hUC-MSCs exosomes(hUC-MSCs-ex)was evaluated by the BCA assay.Concanavalin A and rhIL-2 stimulated umbilical cord blood mononuclear cells (UBMCs) from healthy donor were co-cultured at different concentrations of hUC-MSCs-ex from different cell lines for 72 h.The growth rate of UBMCs was detected by MTT assay.ELISA was used to test the levels of IFN-γ and TNF-α of the supernatants.The UBMCs co-cultured with hUC-MSCs were co-cultured with K562 cells at the ratio of 5∶1.The cytotoxic activity was calculated by MTT assay.Results hUC-MSCs-ex expressed CD9,CD63,CD81and CD44.Different hUC-MSCs lines had different regulating activities on the proliferation,secretion and killing ability of UBMCs.Conclusion hUC-MSCs-ex has heterogeneity of immunomodulatory function on UBMCs.

Intrauterine vertical transmission of HBV via pathway of peripheral blood mono-nuclear cells / 中国免疫学杂志

Objective:To study the HBV infection in peripheral blood mononuclear cells in mediating the role of mother -to-child transmission of hepatitis B virus.Methods: The peripheral blood mononuclear cells ( PBMCs ) in maternal and cord blood mononuclear cells ( CBMCs ) in newborns were conventionally isolated by Ficoll-Hypaque medium.The loads of HBV-DNA in peripheral blood of maternal and cord blood of newborns were both detected by PCR .Results:The clinical data showed that the positive detection rates of HBV-DNA in serum and PBMCs of pregnant women with HBeAg (+) were 100.00%( 25/25 ) and 72.00%( 18/25),and the positive detection rates of HBV-DNA in the neonatal umbilical cord blood serum and CBMCs were 60.00%(15/25) and 44.00%(11/25),respectively.There were significantly difference between HBeAg (+) and HBeAg(-) in the pregnant women (P<0.05 ).The positive detection rates of HBV-DNA in neonatal umbilical cord blood serum and CBMCs were higher in the group with high HBV loads (more than 106copies/ml) in PBMCs than those of low HBV loading group (102-103copies/ml).The significantly difference was explored between the two groups.Conclusion: Mononuclear cells can not only be infected by HBV , but also play a critical role in the intrauterine vertical transmission of HBV via the pathway transmitted from PBMCs in pregnant women to CBMCs in newborns.

Effects of intravenous transplantation of human umbilical cord blood mononuclear cells combined compound Danshen dripping pills on the microenvironment and apoptosis in the myocardium of the rabbits with acute myocardial infarction / 中南大学学报(医学版)

Objective:To explore the effects of compound Danshen dripping pills (CDDP) and CDDP combined with transplantation of human umbilical cord blood cells (HUMNCs) on the inlfammatory response, oxidative stress, myocardial cell apoptosis and cardiac function, and also to investigate the possible mechanisms of the combined therapy in the acute myocardial infarction (AMI). Methods:Rabbit model of AMI successfully established by ligation of the letf anterior coronary artery (LAD). Forty rabbits were randomly divided into 4 groups (n=10 per group):a control group, injected with 0.5 mL of saline in 24 h atfer AMI and then gavaged with 5 mL of saline daily;a CDDP group, injected with saline 0.5 mL atfer AMI and then gavaged with CDDP (270 mg/d) daily;a transplantation group, injected with 0.5 mL of saline contained 3 × 107 HUCBMCs [labeled with green fluorescent protein (GFP)] and then gavaged with 5 mL of saline daily;a combined group, injected with 0.5 mL of saline contained 3 × 107 HUCBMCs (labeled with GFP) and then gavaged with CDDP (270 mg/d) daily. Cardiac function index such as left ventricular fractional shorting (LVFS) and ejection fraction(LVEF) were measured by echocardiography;the pathological changes were observed by HE staining and the white blood cells in the myocardium were determined by light microscopy. hTe superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in myocardium were detected by nitrotetrazolium blue chloride (NBT) and thiobarbituric acid colorimetric measurement respectively. hTe number of transplanted cells in the myocardium was examined by GFP positive cells counted with lfuorescence microscopy. Results:1) Compared with the control group (at 1 or 4 week), LVEF and LVFS were signiifcant improved in the CDDP group, the transplantation group and the combined groups (all P Conclusion:The intravenous transplantation of HUMNCs combined with the CDDP in the treatment of rabbits with AMI could increase the survival rate of transplanted cells and inhibit the myocardial cell apoptosis, therefore improve the heart function. hTe possible mechanism of the combined treatment may be involved in the inhibition of the inlfammatory response and oxidative stress in the myocardium following AMI.

Human Umbilical Cord Blood Mononuclear Cell Transplantation in Rats with Intrinsic Sphincter Deficiency

To evaluate the effectiveness of the human umbilical cord blood (HUCB) transplantation for the treatment of intrinsic sphincter deficiency (ISD), we analyzed the short term effects of HUCB mononuclear cell transplantation in rats with induced-ISD. ISD was induced in rats by electro-cauterization of periurethral soft tissue with HUCB mononuclear cell injection after 1 week. The sphincter function measured by mean leak point pressure was significantly improved in the experimental group compared to the control group at 4 weeks. (91.75+/-18.99 mmHg vs. 65.02+/-22.09 mmHg, P=0.001). Histologically, the sphincter muscle was restored without damage while in the control group it appeared markedly disrupted with atrophic muscle layers and collagen deposit. We identified injected HUCB cells in the tissue sections by Di-I signal and Prussian blue staining. HUCB mononuclear cell injection significantly improved urethral sphincter function, suggesting its potential efficacy in the treatment of ISD.

Inducement of Dendritic Cells Derived from Cord Blood Mononuclear Cells and CTLs Antitumor Immunity in Vitro / 昆明医科大学学报

Objective To investigate the way of inducing dendritic cells from precursor in human cord blood and its role in antitumor immunity.Methods Cord blood was collected under sterile condition and the cord blood mononuclear cells were separated by centrifugal in density gradient.CBMCs were cultured with GM-CSF+IL-4+TNF-?and cell phenotype was analyzed with CD1a、CD83 antibody by using indirect immunofluorescence assays.The effects of DCs pulsed with tumoral antigens on cytotoxic T lymphocytes(CTLs) inducement and growth inhibition of YTMLC cells were assayed.Results Our results indicated that DCs precursors in human cord blood can be induced to differentiate in the medium containing GM-CSF、IL-4 and TNF-?.The cells with typical morphological properties of DCs were observed at the 7th day.At that time,(20.8?1.62)%CD1a+ cells were obtained.After incubation with tumor cytolysis antigen,the DCs can activate the CTLs to become tumor specialized CTLs,which had shown significantly inhibition on growth of YTMLC tumor cell line.Conclusion The precursors in human cord blood can be induced to functional DCs which activate T lymphocyte to become tumor specialized CTLs.

Human cord blood mononuclear cells in promotion to axonal regeneration of injured spinal cord / 中国矫形外科杂志

[Objective]To discuss axonal regeneration and functional recovery after rat's spinal cord injury with transplantation of human cord blood mononuclear cells(HCMNCs).[Method]Thirty injured spinal cord models of Wistar rat were made with Impactor ModelⅡat T10 and then divided into 3 groups randomly and evenly(DMEM control,HCMNCs grafted 3 days post injury and HCMNCs grafted 14 days post injury).Hindlimb functional recovery of rats in each group was evaluated by BBB locomotor functional scale.HE,immunohistochemistry staining,and BDA anterograde tracing were used to observe the axonal regeneration in the lesion site.[Result]Compared with control group,grafted HCMNCs exerted larger effect on promoting nerve regeneration and functional recovery.Rats in the group of HCMNCs grafted 14 days post-injury had better functional recovery than those in HCMNCs grafted 3 days post-injury group.Statistic difference existed among three groups(P

Mixed Mononuclear Cell Culture of Cord Blood and Adult Peripheral Blood / 대한소아혈액종양학회지

PURPOSE: Umbilical cord blood as the source of hematopoietic stem cells has several advantages over bone marrow cells for transplant purpose, but the limitation inherent in the cord blood is its limited number of hematopoietic stem cells. The responses of cord blood mononuclear cells (MNC) which were stimulated by unrelated cord blood MNC or adult peripheral blood MNC to investigate the possibility of unrelated, HLA-mismatched multiple cord blood transplantation were evaluated. METHODS: Nineteen cord blood and 7 adult peripheral blood samples were used. Each of the MNC from cord blood and adult peripheral blood was cultured before and after 2,000 cGy irradiation. Mixed MNC cultures with cord blood and irradiated different cord blood or irradiated adult peripheral blood were performed. RESULTS: The mean percentages of MNC counts compared to day 0 of day 2 and day 7 cultures of mixed MNC cultures with cord blood and irradiated different cord blood were 92.5 55.8%, 57.4 51.0% and those of mixed MNC cultures with cord blood and irradiated adult peripheral blood were 143.4 132.7%, 113.4 95.1%, respectively. The difference of the cell count changes in 2 types of MNC cultures was statistically significant (P<0.05). CONCLUSION: The proliferative responses of cord blood MNC to different cord blood MNC were significantly lower than those to adult peripheral blood MNC. The study for the investigation of clinical applicability of cord blood transplantation from multiple unrelated donors to overcome the limitation of the cell dose of cord blood transplantation and the shortage of related or unrelated HLA-matched donors is needed.