Objective To study the immunomodulatory functions of adult human bone marrow-derived mesenchymal stem cells in vitro.Methods Adult human BMMSCs were separated with Percoll((1.073) g/ml) and cultured in Dulbecco's Modified Eagle's Medium with low glucose containing 10% fetal calf serum.The purity of BMMSCs was identified with the spindle-fibroblastic morphology by microphotograph and the phenotypes were tested by fluorescence-activated cell sorter(FACS).BMMSCs were plated in 96-well plates.After treated with mitomycin,BMMSCs were co-cultured for 72 h with allogeneic lymphocytes isolated from peripheral blood.Effects of MSCs on PHA induced lymphocytes transformation were investigated.The proliferation of lymphocytes was measured by MTT method.Results Peripheral blood lymphocytes proliferation was suppressed by BMMSCs and the inhibition ratio was (60.68%)(P

Objective To approach the dynamic changes of CD~+_ 34 cells and T lymphocyte subsets and best time of peripheral blood stem cell harvest when using rHuG-CSF for peripheral blood stem cell mobilization in donors and patients.Methods From 2001 to 2002 12 donors and 16 patients in Lanzhou General Hospital who received chemotherapy 7 days ago were injected rHuG-CSF 300 ?g/d for mobilization peripheral blood stem cells,and flow cytometry were used to detect the changes of CD~+_ 34 cells and T lymphocyte subsets for 5 days.Results (1)Before using rHuG-CSF,there was obvious difference between patients and donors in bone marrow CD~+_ 34 cells(P

Objective To investigate the effect of immunocompetent cells and immune regulator on the apoptosis of human mesenchymal stem cells ( MSCs) and on mRNA expression of heme oxygenase-1. Methods MSCs were cultured by density gradient centrifugation and then identified by flow cytometry. RT-PCR was used to detect HO-1 mRNA expression and flow cytometry was used to analyze cell apoptosis after the stimulation of IFN-? and PHA-activated T cells. Results The mRNA expression of Heme oxygenase-1 was observed in MSCs and decreased after the stimulation of IFN-? and activated T cells. IFN-?,znpp-Ⅸ and combined these two caused obvious cell apoptosis in MSCs,with an apoptotic rate of ( 56. 50 ? 0. 16) % ,( 56. 85 ? 2. 27) % ,and ( 82. 53 ? 2. 65) % respectively. All of them had a significant difference compared with the normal MSCs [( 7. 56 ? 1. 43) % ,P

Objective To explore the feasibility and safety of cotransplantation of autologous bone marrow-derived mesenehymal stem cells (MSCs) and peripheral blood stem cells in hematological malignant diseases and to observe its effect on hematopoietic reconstruction after cotransplantation. Methods Adult human MSCs were isolated from the healthy bone marrow of the patient himself with Percoll (1. 073 g/ml) and cultured in Dulbecco's modified Eagle's medium with low glucose containing 10% AB type human serum. After conditioning regimen of high-dose chemotherapy and radiotherapy, cotransplantation of autologous bone marrow-derived MSCs and peripheral blood stem cells was done in five patients with hematological malignant diseases. Results The process of the infusion was safe and there were no adverse reactions or other toxicities related to the infustion of MSCs. The median time to achieve neutrophil counts greater than 0. 5 × 109/L was 9.4 days ( ranging from 8 to 11 days) after cotransplantation and platelet counts greater than 20 × 109/L 12. 2 days (ranging from 10 to 14 days). Conclusion Cotransplantation of autologous bone marrow-derived MSCs and peripheral blood stem cells in hematological malignant diseases is feasible and safe. The rapid hematopoietic reconstruction after cotransplantation shows that MSCs have an effect on hematopeiesis, but the mechanism is still to be investigated.

Objective To approach curative effect of using DLI +IL-2 as immunobiotherapy after Mixed-HSCT in acute myelogenous leukemia.MethodAfter times of chemotherapy,8 cases of patients with acute myelogenous leukemia received Mixed-HSCT,then were treated with DLI +IL-2 for 2-7 times.Observed clinical effect for 1 to 5 years.Result DFS in 8 cases of patients with acute myelogenous leukemia received Mixed-HSCT and treated with DLI +IL-2 for 2-7 times were 62.5%.There were no GVHD.Conclusion Immuno-biotherapy with DLI +IL-2 after Mixed-HSCT in patients of acute myelogenous leukemia may be a method to increase DFS efficiently.

BACKGROUND:The incidence rate of peripheral T cel lymphoma is high in Asia, and peripheral T cel lymphoma is aggressive with generaly poor prognosis. However, there is no standard treatment strategy. OBJECTIVE:To retrospectively analyze the therapeutic effect of autologous hematopoietic stem cel transplantation on peripheral T cel lymphoma as wel as relevant toxic and side effects. METHODS:A retrospective review was conducted in 35 patients with peripheral T cel lymphoma who underwent autologous hematopoietic stem cel transplantation from March 2003 to April 2014, including 22 cases of extranodal NK/T-cel lymphoma (nasal type), 1 case of angioimmunoblastic T-cel lymphoma, 8 cases of peripheral T cel lymphoma (non-specific), 3 cases of ALK-positive anaplastic large cel lymphoma, and 1 case of ALK-negative anaplastic large cel lymphoma. Al of 35 patients were classified pathologicaly according to WHO pathological type in 2001 and 2008, and received the high-dose chemotherapy with vincristine, cytarabine, etoposide, mitoxantrone, semustine, cyclophosphamide, and total body irradiation. RESULTS AND CONCLUSION: After a median folow-up of 54 (9-120) months, the probabilities of overal survival and disease-free survival after transplantation were 80% (n=28) and 71% (n=25), respectively. Eight cases (23%) relapsed after transplantation, seven of which died. It was safe with mild and moderate transplantation related side-effects on opportunistic infections, oral cavity mucosa and bladder responses and so on, and there were no severe, life-threatening late complications. Autologous hematopoietic stem cel transplantation may be an effective and safe treatment for peripheral T cel lymphoma, and there is a better benefit in peripheral T cel lymphoma patients with first complete remission.

Objective To explore the dynamic changes of CD34 + cells and T lymphocyte subsets and best time of harvesting peripheral blood stem cell when G-CSF was used for peripheral blood stem cell mobilization in donors and patients. Methods A total of 12 donors and 16 patients who received chemotherapy for 7 d were injected G-CSF of 300 ?g/d to mobilize peripheral blood stem cells for 5 d and flow cytometry were used to detect the changes of CD34 + cells and T lymphocyte subsets everyday for 5 d. Results ① Before G-CSF treatment, there were obvious differences in bone marrow CD34 + cells between patients and donors (P

BACKGROUND:Cobalt chloride (CoCl2) may promote the proliferation of human umbilical cord-derived mesenchymal stem cels (hUC-MSCs) in a time- and concentration-dependent manner, and meanwhile, CoCl2 can regulate the expression of genes and proteins in hUC-MSCs. OBJECTIVE:To explore the effects of CoCl2 induced-hypoxia on the proliferation of hUC-MSCs and gene and protein expressions in hUC-MSCs, thereby establishing an effective method for MSCs culture and amplificationin vitro. METHODS: hUC-MSCs were extracted using tissue explant method. Under hypoxia conditions induced by CoCl2 (0, 100, 150, 200, 250 μmol/L) for different periods (0, 1, 2, 3, 4 days), flow cytometry was used to identify cel surface-associated antigens; cel counting kit-8 was used to detect cel proliferation; RT-PCR was used to determine levels of hypoxia inducible factor-1α, inducible nitric oxide synthase, stromal cel-derived factor-1, interleukin-6, transforming growth factor-β mRNA; western blot assay was used to detect protein expression of hypoxia inducible factor-1α. RESULTS AND CONCLUSION:The cels were positive for CD29, CD73, CD90, CD105, while negative for CD31, CD14, CD34, CD45, CD11b, HLA-DR. Moreover, the antigen expression was not affected by CoCl2 induced-hypoxia. CoCl2 induced-chemical hypoxia could promote the proliferation of hUC-MSCs in a time- and concentration-dependent manner. RT-PCR results showed thatunder hypoxia, hypoxia inducible factor 1α, inducible nitric oxide synthase and stromal cel-derived factor-1 mRNA expressions were significantly up-regulated, but interleukin-6 and transforming growth factor-β mRNA expressions were down-regulated significantly (P < 0.05). Additionaly, the protein expression of hypoxia inducible factor 1α was increased under hypoxia conditions. These findings indicate that CoCl2 induced-hypoxia environment may promote the proliferation of hUC-MSCs and the optimal concentration of CoCl2 is 200μmol/L. However, a higher concentration of CoCl2 (≥ 250μmol/L) inhibits the proliferation of hUC-MSCs, and the mechanism may be related to the increase of hypoxia inducible factor-1α at protein and mRNA levels.

Objective To investigate the curative effect of allogeneic hematopoietic stem cell transplantation (allo-HSCT) and the method of the pretreatment choice on myelodysplastic syndrome (MDS).Methods 13 cases who were undergoing allo-HSCT including 10 HLA-matched, 2 HLA partially matched,and one umbilical cord blood transplanted patients were enrolled in this study. The infusion number of MNC and CD+34 cells is 6.92 (2.65-21.33)×108/kg and 4.47 (1.49-10.22) ×106/kg, respectively. Of the 13 cases,5 adopted TBI, fludarabine(Flud), and cyclophosphamide(Cy) pretreatment, 3 chose BU/Cy pretreatment, and 3 chose TBI and Cy pretreatment, 2 chose Ara-C+BU+Cy+VM26 pretreatment. In order to prevent GVHD, 2cases of HLA partially matched were treated with ATG, cyclosporin A (CsA), MTX and MMF aftertransplantation, while the others were treated with CsA and MTX only. Results 9 out of 13 cases achieved hematopoietic reconstruction completely. 4 died of complications. Conclusion The allogeneic hematopoietic stem cell transplantation is an efficient regimen to cure MDS. The pretreatment should adopt the individuation.

Objective To observe the effects of low dose irradiation (LDR) on proliferation,adipogenesis and osteogenic potential of human umbilical cord mesenchymal stem cells (hucMSCs).Methods hucMSCs were isolated from Wharton's jelly tissue of human umbilical cord by modified tissuepiece inoculation,and flow cytometry was used to detect the expression of specific marker in the hucMSCs.The hucMSCs were randomly divided into two groups:irradiation group undergoing irradiation with the doses 50,100,or 200 mGy respectively,and control group without irradiation.MTT method was applied to evaluate the proliferation of the hucMSCs at different time points with various doses irradiation.The third passage hucMSCs were randomly divided into two groups:irradiation group undergoing low dose irradiation of 200 mGy,and control group without irradiation,and then underwent induction by adipocytic and oesteocytic differentiation induction fluids respectively so as to differentiate into adipocytes and osteoblasts.Oil red O staining was used to detect the activity of alkaline phophatase (ALP),and RT-PCR was used to detect the mRNA expression of core binding factor alpha 1 in human osteoblast.Results After 9-12days,fibroblasts began to swim out of the tissue piece with a confluence rate of 80% 2 weeks later.Within 7 days the absorption values of the hucMSCs undergoing different irradiation doses 2,3,4,5,and 6 days later were all significantly higher than those of the control group(F = 159.17,448.81,265.15,183.93,and 181.83 ,all P <0.01),with the proliferation rates of the 100 mGy subgroup being the highest.After being induced liquid,vacuoles were observed in the irradiated group 2 days later.21 days later,the adipogenic rates of irradiated group was significantly higher than that of the control group (t = 28.25,P <0.01).The ALP activity increased in the irradiated group compared with control group (t=16.87,P <0.01) .The expression level of Cbf-α1 mRNA was up-regulated obviously (t = 14.16,P＜0.01).Conclusions LDR promotes the proliferation of hucMSCs,and accelerates the hucMSCs' differentiation into adipocytes and osteoblasts.