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Qian Y.-X.,Shanghai Cord Blood Bank | Shu Q.,Changning District Maternal and Child Health Hospital | Cai H.-X.,Changning District Maternal and Child Health Hospital | Guo J.,Shanghai Sixth Peoples Hospital | And 2 more authors.
Journal of Clinical Rehabilitative Tissue Engineering Research | Year: 2011

BACKGROUND: Mesenchymal stem cells are the stem cells that possess the capability for self-renewal and multi-directional differentiation. Umbilical cord is the tissue outside the embryos and would be fallen off after parturition. In addition, it has wide source and no ethical restriction, so it is promising to be the first choice for mesenchymal stem cells. OBJECTIVE: To detect the surface markers CD29, CD44, CD49e, CD73, CD90, CD34, CD45, and CD271 of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) prior to and after cryopreservation and resuscitation. METHODS: After isolation and culture, morphology of the primary, P4 and P8 hUCMSCs was observed prior to cryopreservation and after resuscitation. Surface markers CD29, CD44, CD49e, CD73, CD90, CD34, CD45, and CD271 of primary, P4, and P8 hUCMSCs were detected through the use of flow cytometry prior to cryopreservation and after resuscitation RESULTS AND CONCLUSION: hUCMSCs prior to cryopreservation and hUCMSCs of different passages after resuscitation present the same phenotype, i.e., positive for CD29, CD44, CD49e, CD73, and CD90, and negative for CD34, CD45, CD271. These findings suggest that primary hUCMSCs do not present changes in surface markers after cryopreservation and resuscitation.

Fan D.,Shanghai JiaoTong University | Zhou X.,Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education | Li Z.,Ruijin Hospital | Li Z.-Q.,Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education | And 13 more authors.
Oncogene | Year: 2015

Leukemic lymphoblasts within different immunophenotypic populations possess stem cell properties. However, whether or not the self-renewal program is retained from stem cells or conferred on progenitors by leukemogenic molecules remains unknown. We have addressed the issue in the context of TEL-AML1-associated acute lymphoblastic leukemia (ALL) by profiling a refined program edited from genes essential for self-renewal of hematopoietic stem cells and B-cell development. Bioinformatic analysis shows that ALL populations are loosely clustered and close to the normal population that contains stem and primitive progenitor cells. This finding indicates that immunophenotypes do not reflect maturation stages in ALL and that the self-renewal program may be retained from stem cells. Results of assessing 'first hit' function of TEL-AML1 in different populations of normal cells demonstrate the molecular model. Therefore, the current study shows a leukemogenic scenario of human ALL in which programs of stem cells are sustained in distinct fractions by leukemogenic mutations.

Fan H.,East China Normal University | Fan H.,Shanghai Blood Center | Yang J.,Shanghai Blood Center | Hao J.,East China Normal University | And 7 more authors.
Immunology | Year: 2012

In this study, we expanded regulatory T cells (Tregs) ex vivo from CD4 +CD25 + T cells from cord blood (CB) and CD4 +CD25 +CD127 - T cells from adult peripheral blood (APB) and compared the suppressive functions of the newly generated Tregs. The Tregs from CB and APB were expanded either in two cycles with a polyclonal stimulus or in two cycles with an alloantigen stimulus in the first cycle and a polyclonal stimulus in the second cycle. Cell yield after Treg expansion with polyclonal stimulation was greater than that of Tregs expanded with combined alloantigen and polyclonal stimulation. The expanded Tregs expressed high levels of Foxp3, CD39 and cytotoxic T-lymphocyte antigen-4 and low levels of CD127, interleukin-2 and interferon-γ. After two cycles of expansion, the CB Tregs maintained expression of the GARP gene and showed greater suppressive function than APB Tregs. The CB Tregs that were expanded with two cycles of polyclonal stimulation suppressed not only the polyclonal antigen-driven responder T (T resp) cell proliferation but also the HLA mismatched dendritic cell-driven T resp cell proliferation. When CB and APB Tregs were expanded with a primary alloantigen stimulus followed by a secondary polyclonal stimulus, the Tregs showed a potent, antigen-specific suppressive capacity. The Tregs expanded with two cycles of polyclonal stimulation from both CB and APB alleviated acute graft-versus-host disease symptoms and prolonged survival in a murine model of graft-versus-host disease. In conclusion, CB Tregs expanded with two cycles of polyclonal stimulation had a stronger immunosuppressive function than APB Tregs. It is feasible to obtain human functional alloantigen-specific Tregs expanded ex vivo from CB and APB in large numbers. © 2012 The Authors. Immunology © 2012 Blackwell Publishing Ltd.

Chang H.-R.,Soochow University of China | Cheng Z.,Soochow University of China | Chen G.-H.,Soochow University of China | Sun A.-N.,Soochow University of China | And 13 more authors.
Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology | Year: 2014

This study was purposed to investigate the engraftment, graft-versus-host disease (GVHD), transplantation related mortality (TRM), relapse and survival in hematologic patients received unrelated umbilical cord blood transplantation (UCBT). A total of 25 patients with hematological disease underwent UCBT, including 8 pediatric and 17 young adult patients. Among them 3 cases received single unit of UCBT and 22 cases received double units of UCBT. For donor/recipients human leukocyte antigen (HLA) matching: HLA 6/6 loci matched in 9 cases, HLA 4-5/6 loci matched in 16 cases. There were 19 patients with hematologic malignancies, including 3 cases in the period of disease progression and 6 cases of non-hematologic malignancies. Conditioning regimens were TBI/Cy ± Flu ± ATG or BuCy ± Flu ± ATG for 21 patients and Cy+Flu+ATG for 4 patients. For prophylaxis of acute graft-versus-host disease (aGVHD) the regimen of cyclosporine (CsA) as dominant drug was used. The results showed that among 16 patients (80.0%) achieved engraftment, 20 patients survived for more than 42 d after transplantation. The cumulative neutrophil recovery rate on day 42 after transplant was 64.0%, with a median time of 17.0 d;the cumulative platelet recovery rate on day 100 after transplant was 60.0 %, with a median time of 35.0 d. The cumulative rate of grade II-IV and III-IV aGVHD after transplantation 100 d was 44.0% and 30.7%, respectively. Until the end of the follow-up, the cumulative rate of TRM was 54.3%. For all the patients, overall survival rate was 42.7%. Out of 17 evaluable patients with hematologic malignancies 7 cases (41.2%) survived to date, and only 1 case relapsed, so event-free survival rate was 35.3%. Out of 5 evaluable patients with non-hematologic malignancies, 4 patients survived and 2 patients were in stable engraftment state, 2 cases with autologous hematopoietic recovery. Among 3 cases of hematologic malignancies at advanced stage, only 1 case survived to date. It is concluded that HLA-4-6/6 loci matched UCBT is an effective option to treat hematological diseases. Double cord blood transplantation (dUCBT) can overcome the disadvantage of insufficient cells of single cord blood UCBT to treat overweight children and adult.

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