Spleen regeneration after subcutaneous heterotopic autotransplantation in a mouse model

BACKGROUND: Splenectomy may lead to severe postoperative complications, including sepsis and cancers. A possible solution to this problem is heterotopic autotransplantation of the spleen. Splenic autografts rapidly restore the regular splenic microanatomy in model animals. However, the functional competence of such regenerated autografts in terms of lympho- and hematopoietic capacity remains uncertain. Therefore, this study aimed to monitor the dynamics of B and T lymphocyte populations, the monocyte-macrophage system, and megakaryocytopoiesis in murine splenic autografts. METHODS: The model of subcutaneous splenic engraftment was implemented in C57Bl male mice. Cell sources of functional recovery were studied using heterotopic transplantations from B10-GFP donors to C57Bl recipients. The cellular composition dynamics were studied by immunohistochemistry and flow cytometry. Expression of regulatory genes at mRNA and protein levels was assessed by real-time PCR and Western blot, respectively. RESULTS: Characteristic splenic architecture is restored within 30 days post-transplantation, consistent with other studies. The monocyte-macrophage system, megakaryocytes, and B lymphocytes show the highest rates, whereas the functional recovery of T cells takes longer. Cross-strain splenic engraftments using B10-GFP donors indicate the recipient-derived cell sources of the recovery. Transplantations of scaffolds populated with splenic stromal cells or without them afforded no restoration of the characteristic splenic architecture. CONCLUSIONS: Allogeneic subcutaneous transplantation of splenic fragments in a mouse model leads to their structural recovery within 30 days, with full reconstitution of the monocyte-macrophage, megakaryocyte and B lymphocyte populations. The circulating hematopoietic cells provide the likely source for the cell composition recovery.

Reconstitución de células natural killer después del trasplante de células progenitoras hematopoyéticas en niños / Natural killer cell reconstitution after hematopoietic stem-cell transplantation in children

Resumen Introducción: Después de un trasplante de células progenitoras hematopoyéticas (TCPH), la reconstitución de las células natural killer (NK) es la principal barrera contra las infecciones virales. Objetivo: Determinar que el conocimiento sobre la cinética de la reconstitución de las células NK posterior al TCPH contribuye a un eficiente monitoreo del trasplante, lo que incrementa la posibilidad de éxito de este. Método: Se incluyeron 21 pacientes sometidos a TCPH, así como un grupo control de individuos clínicamente sanos. En diferentes momentos después del trasplante (intervalo de 21 a 670 días), mediante citometría de flujo se cuantificaron las células NK CD3− CD16+ CD56+ en muestras de sangre periférica. Resultados: La recuperación de las células NK ocurre entre los tres y seis meses y entre los 10 y 12 meses postrasplante; su número fue significativamente menor (en comparación con el grupo control) en el tiempo restante del monitoreo. Conclusiones: El primer periodo de recuperación de las células NK ocurre entre los tres y seis meses posteriores al trasplante. La reconstitución es transitoria y el número de células NK varía en los primeros años.

Abstract Introduction: After hematopoietic stem cell transplantation (HSCT), natural killer (NK) cells reconstitution is the main barrier against viral infections. Objective: To determine that the knowledge on the kinetics of NK cell reconstitution after HSCT contributes to transplant efficient monitoring, which increases the possibility of its success. Method: Twenty-one patients undergoing HSCT were included, as well as a control group of clinically healthy individuals. At different time points after transplantation (range of 21 to 670 days), CD3- CD16+ CD56+ NK cells were quantified by flow cytometry in peripheral blood samples. Results: NK cell recovery occurs at three to six months and 10 to 12 months post-transplantation; their number was significantly lower (in comparison with the control group) in the rest of the monitoring time. Conclusions: The first period of NK cell recovery occurs between three and six months after transplantation. Reconstitution is transient and the number of NK cells varies in the first years.

Developmental relationship between mesenchymal stem/progenitor cells and hematopoietic cells during human embryogenesis / 中华微生物学和免疫学杂志

Objective@#To analyze the developmental relationship between mesenchymal stem/progenitor cells (MSPCs) and hematopoietic cells during human embryogenesis.@*Methods@#Aborted embryos at different developmental stages were used in this study after medical abortion. Embryonic blood tissues were isolated and digested into single cells. These single cells were plated in semisolid medium in favor of the differentiation of colony-forming cell with high proliferative potential (HPP-CFC) and incubated for 10 to 14 d. Individual colonies with diameter more than 0.5 mm were picked and replated in liquid medium. Fibroblastic adherent cells appeared in the replated colonies were cultured for cell proliferation and cytokins expressed on cell surface were identified to analyze whether they had the characteristics of MSPCs.@*Results@#This study summarized the dynamic development of HPP-CFCs and other hematopoietic progenitor cells in different tissues including aorta-gonad-mesonephros (AGM) region, yolk sac and embryonic liver. From the 28-somite stage, a proportion of HPP-CFCs in AGM region could give rise to adherent fibroblastic cells in addition to hematopoietic cells. The adherent cells harbored the differentiation potential of MSPCs and could inhibit the proliferation of T cells in lymphocyte transformation test.@*Conclusions@#This study suggests some prehematopoietic precursors in AGM region can give rise to both hematopoietic progenitors and MSPCs during human embryogenesis.

Developmental relationship between mesenchymal stem/progenitor cells and hematopoietic cells dur-ing human embryogenesis / 中华微生物学和免疫学杂志

Objective To analyze the developmental relationship between mesenchymal stem/pro-genitor cells (MSPCs) and hematopoietic cells during human embryogenesis. Methods Aborted embryos at different developmental stages were used in this study after medical abortion. Embryonic blood tissues were isolated and digested into single cells. These single cells were plated in semisolid medium in favor of the dif-ferentiation of colony-forming cell with high proliferative potential ( HPP-CFC) and incubated for 10 to 14 d. Individual colonies with diameter more than 0. 5 mm were picked and replated in liquid medium. Fibroblastic adherent cells appeared in the replated colonies were cultured for cell proliferation and cytokins expressed on cell surface were identified to analyze whether they had the characteristics of MSPCs. Results This study summarized the dynamic development of HPP-CFCs and other hematopoietic progenitor cells in different tis-sues including aorta-gonad-mesonephros ( AGM) region, yolk sac and embryonic liver. From the 28-somite stage, a proportion of HPP-CFCs in AGM region could give rise to adherent fibroblastic cells in addition to hematopoietic cells. The adherent cells harbored the differentiation potential of MSPCs and could inhibit the proliferation of T cells in lymphocyte transformation test. Conclusions This study suggests some prehemato-poietic precursors in AGM region can give rise to both hematopoietic progenitors and MSPCs during human embryogenesis.

Primary intraosseous hematopoietic pseudotumor: clinicopathological analysis and 9-year follow-up of 3 cases / 南方医科大学学报

We analyzed the clinicopathological data of 3 cases of primary intraosseous hematopoietic pseudotumor (IHPT), which had been previously misdiagnosed as malignancies or metastases both clinically and pathologically. Two of the patients received close follow-up for 132 and 100 months, and one patient was lost to follow-up, and the tumors were confirmed to be benign in all the 3 cases. IHPT is a rare benign intraosseous solid lesion consisting of tissues resembling normal hematopoietic tissue, and can be easily misdiagnosed as malignancy. Understanding the clinicopathological features and the outcomes of the disease can facilitate the clinical decisions on individualized diagnosis and therapeutic regimens.

Hipertensão pulmonar após transplante autólogo de células-tronco hematopoiéticas em neuroblastoma / Pulmonary hypertension after autologous transplant of stem-hematopoietic cells in neuroblastoma

Objetivo: Relatar o caso de um paciente pediátrico pós-transplante de células-tronco hematopoiéticas (TCTH) que teve o diagnóstico de doença vaso-oclusiva pulmonar (DVOP), assim como a conduta terapêutica adotada e resolução do caso. Método: Menino de 1 ano de idade, com diagnóstico de neuroblastoma IV, que pós- TCTH evoluiu para DVOP. Conclusão: O caso apresentado condiz com os pouquíssimos casos relatados na literatura de hipertensão pulmonar após TCTH. Criança com neoplasia maligna, condicionamento mieloablativo e tempo de aparecimento da HP dentro da média descrita pela literatura. A causa mais provável da HP foi DVOP, uma vez que o paciente apresentou doença veno-oclusiva hepática (DVOH).

Design and implementation of the TRACIA: intracoronary autologous transplant of bone marrow-derived stem cells for acute ST elevation myocardial infarction / Diseño e implementación del estudio TRACIA: Trasplante intracoronario Autológo de Células madre para Infarto Agudo del miocardio con elevación de segmento ST

Objective: To describe the design of a protocol of intracoronary autologous transplant of bone marrow-derived stem cells for acute ST-elevation myocardial infarction (STEMI) and to report the safety of the procedure in the first patients included. Methods: The TRACIA study was implemented following predetermined inclusion and exclusion criteria. The protocol includes procedures such as randomization, bone marrow retrieval, stem cells processing, intracoronary infusion of stem cells in the infarct-related artery, pre-and-post MRI, pre-and-post SPECT with radioisotope ventriculography, and clinical follow-up at 6 months. Results: Eight patients with a diagnosis of acute STEMI and duration of symptoms of <24 hours that were perfused successfully through primary percutaneous coronary intervention (PPCI) with a LVEF of <45% were assigned randomly to two groups (n = 4 each). One group treated with stem cells and the other corresponded to the control group. Neither death, re-infarction, no need for revascularization or thrombosis of the stent were observed at follow-up. Conclusions: The initial experience at the Instituto Nacional de Cardiología Ignacio Chávez in the treatment of acute STEMI by means of autologous transplantation of bone marrow-derived stem cells is encouraging. Implementation was possible in the first eight patients with no complications.

Objetivo: Describir el diseño y la implementación de un protocolo de transplante autólogo intracoronario de células madre derivadas de médula ósea en infarto agudo al miocardio con elevación del ST y reportar la seguridad del procedimiento en los primeros pacientes incluidos. Métodos: El estudio TRACIA se implementó con base en criterios de inclusión y exclusión predeterminados. El protocolo incluye la aleatorización, obtención de médula ósea, procesamiento de células madre, infusión intracoronaria de células madre, RM basal y al seguimiento, SPECT con ventriculografía radioisotópica basal y post-procedimiento, y seguimiento clínico a seis meses. Resultados: Ocho pacientes con diagnóstico de infarto agudo del miocardio con elevación del ST y duración de síntomas <24 horas que fueron reperfundidos exitosamente con angioplastia primaria y con fracción de expulsión <45%, fueron aleatorizados a dos grupos; uno de ellos fue tratado con células madre y el otro grupo permaneció como control. No se observó muerte, re-infarto, necesidad de revascularización o trombosis del Stent durante el seguimiento. Conclusiones: La experiencia inicial en el Instituto Nacional de Cardiología Ignacio Chávez en el tratamiento del infarto agudo del miocardio con elevación del ST mediante trasplante autólogo de células madre derivadas de médula ósea, es alentadora. La implementación sin complicaciones fue posible en los primeros ocho pacientes.

Efectos del uso del factor estimulante de colonias granulocíticas en la obtención de células madre en un grupo de pacientes con enfermedad arterial oclusiva crónica / Effects of the use of granulocyte colony stimulating factor for obtaining stem cells in a group of patients with chronic occlusive arterial disease

Es importante evaluar el perfil del uso del factor estimulante de colonias granulocíticas (fec-g) en pacientes con enfermedad arterial oclusiva crónica (eaoc), mediante el análisis de aspectos como eficacia y seguridad. Se examinaron los datos obtenidos de la cohorte de pacientes con eaoc que asistían regularmente a la clínica de Cirugía Vascular del Hospital Militar Central en Bogotá. El protocolo de movilización de células CD34+ hacia sangre periférica consistió en el uso de FEC-Grh a dosis de 600 mg/día por vía subcutánea (Filgrastim fec-g 300 mg Roche®), repartido en dos dosis diarias, en forma continua durante cinco días. Al realizar la comparación de valores a partir de hemogramas realizados antes y después de la movilización, se demostró incremento significativo en el número de leucocitos así como en la proporción de neutrófilos y basófilos; mientras que las proporciones de monocitos, eosinófilos y linfocitos disminuyeron significativamente. Con respecto al comportamiento de las células CD34+, no se muestra una diferencia significativa en el comportamiento del CD34+ con la edad, así como tampoco con el índice de masa corporal (imc). En lo relacionado con el peso y los niveles de CD34+, se observó que los pacientes que lograron una buena respuesta tenían un peso de 59,7 kg, mientras que los pacientes con regular respuesta, 68,1 kg.

The analyzed aspects such as efficacy and safety are important in the use of Granulocyte Colony Stimulating Factor in patients with chronic occlusive arterial disease were analyzed data from the cohort of patients with eaoc that regularly attended the Clinic for Vascular Surgery of the Hospital Militar Central in Bogotá. The protocol for CD34+ cell mobilization into peripheral blood involved the use of FEC-Grh at a dose of 600 mg/day administered subcutaneously (Filgrastim g-csf 300 mg Roche®) divided into two daily doses, continuously for five days. By comparing the values from blood counts performed before and after mobilization showed increased significant number of leukocytes as well as proportion of neutrophils and basophiles, whereas proportions of monocytes, eosinophils and lymphocytes decreased significantly. About the CD34+ cell behavior, it’s not shown significant difference between the behaviors of CD34+ with age, neither the imc. The analyzed done on the weight and CD34+ levels was observed that patients achieved a good response with a weight of 59.7 kg while 68.1 kg patients with regular response.

Adeno-associated Virus Serotype 2 Mediated Transduction and Expression of The Human β-Globin Gene in Human Early Fetal Liver Hematopoietic Cells / 生物化学与生物物理进展

β-Thalassemia is an inheritance anaemia disease due to the defect in β- globin gene. Gene therapy is considered to be the only method which could cure this disease. Adeno-associated virus type 2 (AAV2) has benn gaining more attention as a vector in human gene therapy for its non pathogenic character and broad host range. Although, the efficacy of recombinant AAV2 (rAAV2) in transducing human hematopoietic stem cells has been investigated by researchers, the results were varied from different laboratory. The view was proposed recently that it may be resulted from helper virus in their packaging system. Respecting this, the packaging system without helper virus was used to produce rAAV2. Human early fetal liver hematopoietic cells not only possess many superior peculiarity compared to hematopoietic cells of bone marrow or cord blood, but also the inherent β-globin gene in the cells is not expressed. Studies on the AAV2 transduction of human fetal liver hematopoietic cells and mediated expression of β- globin gene in vivo were performed and the potential role of AAV2 in β-thalassemia gene therapy was analyzed. The rAAV2 containing a normal human β-globin gene (rAAV2-β-globin) without helper virus contamination were produced. The viral titer, purity and the ability of mediating expression of β-globin gene were detected in vitro. Then, human early fetal liver hematopoietic cells were isolated and were further transducted with the rAAV2-β-globin, followed by transplantation into sublethally irradiated BALB/C nude mice to analyze the β-globin gene expression. The results showed that the high titer and purity of rAAV2-β-globin had the ability of mediating β-globin gene expression in vitro. In 8 recipient BALB/C nude mice, the β-globin gene expression were detected in the 2 mice marrow by RT-PCR. The results suggested that rAAV2 could transduce human fetal liver hematopoietic cells and mediate the β-globin gene expression in BALB/C nude mice, meanwhile the expression level of the gene was still rather low. It is necessary to perform further research on AAV2 biology before applying in β-thalassemia gene therapy.

Adeno-associated Virus Serotype 2 Mediated Transduction and Expression of The Human ?-Globin Gene in Human Early Fetal Liver Hematopoietic Cells / 生物化学与生物物理进展

?-Thalassemia is an inheritance anaemia disease due to the defect in?- globin gene. Gene therapy is considered to be the only method which could cure this disease. Adeno-associated virus type 2 (AAV2) has benn gaining more attention as a vector in human gene therapy for its non pathogenic character and broad host range. Although, the efficacy of recombinant AAV2 (rAAV2) in transducing human hematopoietic stem cells has been investigated by researchers, the results were varied from different laboratory. The view was proposed recently that it may be resulted from helper virus in their packaging system. Respecting this, the packaging system without helper virus was used to produce rAAV2. Human early fetal liver hematopoietic cells not only possess many superior peculiarity compared to hematopoietic cells of bone marrow or cord blood, but also the inherent?-globin gene in the cells is not expressed. Studies on the AAV2 transduction of human fetal liver hematopoietic cells and mediated expression of ?- globin gene in vivo were performed and the potential role of AAV2 in?-thalassemia gene therapy was analyzed. The rAAV2 containing a normal human?-globin gene (rAAV2-?-globin) without helper virus contamination were produced. The viral titer, purity and the ability of mediating expression of?-globin gene were detected in vitro. Then, human early fetal liver hematopoietic cells were isolated and were further transducted with the rAAV2-?-globin, followed by transplantation into sublethally irradiated BALB/C nude mice to analyze the?-globin gene expression. The results showed that the high titer and purity of rAAV2-?-globin had the ability of mediating ?-globin gene expression in vitro. In 8 recipient BALB/C nude mice, the ?-globin gene expression were detected in the 2 mice marrow by RT-PCR. The results suggested that rAAV2 could transduce human fetal liver hematopoietic cells and mediate the ?-globin gene expression in BALB/C nude mice, meanwhile the expression level of the gene was still rather low. It is necessary to perform further research on AAV2 biology before applying in ?-thalassemia gene therapy.

Research on the effects and side effects of the over-dose chemotherapy to VX2 hepatocarcinoma rabbits with mdr1 gene transferring bone marrow transplantation / 重庆医科大学学报

Objective:In the study,multidrug resistance gene 1(mdr1) was transferred into the bone marrow mononuclear cells which were autotransplanted into rabbits with VX2 hepatocarcinoma.In chemotherapy experiment with adriamycin,the bone marrow protection of mdr1 gene,curative effects and side effects to heart of over dosage chemotherapy were observed.Methods: The bone marrow mononuclear cells transferred with mdr1 gene were autotransplanted into rabbits with VX2 hepatocarcinoma.After chemotherapy with adriamycin,the peripheral white blood cells count,the growth and metastasis of VX2 hepatocarcinoma and cardiac function were detected.Meanwhile,the bone marrow protection of mdr1 gene,the curative effects to VX2 hepatocarcinoma and lesions to heart of over-dosage chemotherapy were evaluated.Results:The mdr1 gene was implanted into the bone marrow after the mdr1-transferred bone marrow mononuclear cells had been autotransplanted into rabbits with VX2 hepatocarcinoma.After over-dose chemotherapy with adriamycin,the rabbits in mdr1-transferring group could be survival after treated with 3-fold dosage chemotherapy,and the white blood cell count in peripheral blood was(4.26?1.03)?109/L.Meanwhile,the tumor cells were killed effectively,and the survival time were improved compare to that of control group(P

Effects of Short-Term Incubation of Hematopoietic Cells with Hematopoietic Growth Factors on CXCR4 Expression and Stromal Cell-Derived Factor-1-Mediated Transendothelial Migration / 대한혈액학회지

BACKGROUND: There has been contradictory reports regarding the homing potential of hematopoietic cells briefly exposed to hematopoietic growth factors in vitro. To get a resolution to this controversy, we investigated the effects of short-term growth factor treatment of hematopoietic cells on the expression of CXCR4 and adhesion molecules, and the chemotaxis in response to stromal cell-derived factor-1 (SDF-1), which is widely accepted to play a critical role in bone marrow (BM) homing of hematopoietic stem cells. METHODS: BM and cord blood(CB) CD34+ cells were incubated with various hematopoietic growth factors including IL-1beta, IL-3, IL-6, G-CSF, GM-CSF, stem cell factor (SCF), flk-2 ligand, and thrombopoietin, alone or in combination for up to 48 hours. Before and after the incubation, the expression of CXCR4 and adhesion molecules of CD34+ cells was analyzed using flow cytometry. SDF-1-mediated transmembrane or transendothelial migration of CD34+ cells, cobblestone area-forming cells (CAFCs), and/or long-term culture-initiating cells (LTC-ICs) was measured using Transwell(TM) system. RESULTS: VLA-4 was moderately up-regulated by the incubation of the cells with IL-3 and SCF, and ICAM-1 was slightly up-regulated by IL-1 and IL-3. The expression of L-selectin, PECAM-1 or LFA-1 was not altered by any growth factors. With the incubation of the cells in the absence of growth factors or SDF-1, CXCR4 expression of CD34+ cells was rapidly increased, reaching a plateau at 24 hours. The spontaneous up-regulation was abrogated with the addition of SDF-1. In agreement with the up-regulation of CXCR4, CD34+ cells incubated for 40 hours showed much enhanced chemotaxis in response to SDF-1 compared to non-incubated cells (24.7 3.5% vs. 7.0 1.6%, P=0.01). Any growth factors examined in this study did not alter the CXCR4 expression of CD34+ cells. Neither did growth factors affect the transendothelial migration of LTC-ICs toward bone marrow stromal cells as well as the SDF-1-induced transmembrane chemotaxis of CD34+ cells and CAFCs. CONCLUSION: Short-term treatment of hemo-topoietic cells with hematopoietic growth factors does not alter the expression of CXCR4 or SDF-1-mediated transendothelial chemotaxis.

Functional and morphological changes in mouse hematopoietic system after exposure to ?-ray irradiation combined with microwave / 解放军医学杂志

Objective To explore the patterns and features of biological effects of ?-ray irradiation combined with microwave on the mouse hematopoietic system, and to provide theoretical and experimental basis for understanding the possible mechanism of the bone marrow injury caused by the combined action of microwave and ?-ray irradiation. Methods 216 healthy KM mice were randomly divided into the following four groups: normal control, microwave (S frequency range, 50mW/cm~2), ?-ray irradiation (5.5Gy), microwave combined ?-ray radiation (5.5Gy + 50mW/cm~2). They were sacrificed at 6h, 1, 3, 7, 14, 28, 90 and 180 days after radiation, respectively, then the histological and ultrastructural changes in the bone marrow and the peripheral hemogram were observed. Results Histopathological changes: the bone marrow appeared to be obviously injured either by radiation or microwave exposure, characterized by undergoing four phases, namely apoptosis-necrosis, void, regeneration and recovery phase. However, the pathological changes were more obvious and the recovery was slower in microwave combined ?-ray radiation group. Peripheral hemogram: the numbers of leucocytes, erythrocytes and platelets, and the content of hemoglobin decreased in both ?-ray irradiation group and microwave combined ?-ray radiation group, and the decrease in microwave combined radiation group was more remarkable. Ultrastructure: the bone marrow hematopoietic cells underwent obvious degeneration, apoptosis and necrosis in microwave combined ?-ray radiation group especially at 6 hours after radiation. Conclusion ?-ray combined with microwave could induce hematopoietic dysfunction and pathomorphological changes in hematopoietic organ, which were mainly caused by ?-ray, and the changes were aggravated.

Ex Vivo Expansion of Primitive Hematopoietic Cells by Using Hematopoietic Growth Factors / 대한소아혈액종양학회지

PURPOSE: If primitive hematopoietic cells expanded ex vivo can be used in stem cell transplantation, duration for hematologic recovery will be shortened. In this study, CD34 cells were cultured with various hematopoietic growth factors which are known to stimulate proliferation of primitive hematopoietic cells. METHODS: CD34 cells isolated from cord blood were cultured and expanded ex vivo with IL-3, stem cell factor (SCF), flt-3 ligand (FL), thrombopoietin (TPO), granulocyte-colony stimulating factor (G-CSF). To find optimal combination of growth factors, CD34 cells were cultured for 9 days with various combinations of growth factors. To find optimal duration of culture, CD34 cells were cultured for 5, 7, 9, 12 days. To evaluate expansion of primitive hematopoietic cells, the number of CD34 cells, colony forming cells and long-term culture-initiating cells (LTC-IC) were counted by flow cytometry, colony forming cell assay and limiting dilution assay respectively. RESULTS: Primitive hematopoietic cells were successfully expanded from CD34 cells of cord blood. Maximal expansion of LTC-IC and CFU- GEMM were 2.58 and 2.37 fold respectively after 9 days of culture, and were obtained with the combination of IL-3 SCF FL TPO. When CD34 cells were cultured for 5, 7, 9, 12 days with the combination of IL-3 SCF FL TPO, expansion of LTC-IC was maximal (2.95 fold) after 9 days culture. After reaching maximal expansion of LTC-IC, the number of nucleated cells increased, but that of primitive hematopoietic cells decreased. CONCLUSION: Primitive hematopoietic cells can be successfully expanded ex vivo by using hematopoietic growth factors. Duration for hematologic recovery after stem cell transplantation will be shortened by using primitive hematopoietic cells expanded ex vivo.

The stimulative effects of cord blood serum on hematopoietic cells in vitro / 中国免疫学杂志

Objective:To study the effects of human CBS(Cord Blood Serum) on hematopoietic cells culture in vitro.Methods:CBS and/or different combinations of cytokines were added to the culture system derived from human bone marrow mononuclear cells.Colony forming capacity of colony forming unit-granulocyte/ monocyte (CFU-GM)?colony forming unit-granulocyte/ erythrocyte/ megakaryocyte/ monocyte (CFU-GEMM)?cobble-stone area forming cells (CAFC)?long-term culture initiating cells (LTC-IC) were observed.Results:CBS had certain cytokine-like stimulating activity that could promote the proliferation and differentiation of hematopoietic cells of bone marrow in vitro. By comparison with cytokines, its GM-CSF-like activity was equal to 65.6 ?g/L rhGM-CSF, and its IL-3-like activity was equal to 2.9 ?g/L rhIL-3.Conclusion:CBS contains stimulating activities for proliferating and differentiating of hematopoietic cells. CBS alone can maintain hematopoietic cells culture in vitro. This study showed the prospect of clinical application of CBS.

TC-1 Stromal Cells Enhance Retro viral-Mediated Gene Transfer into Murine Marrow Hematopoietic Cells / 中国肿瘤生物治疗杂志

Objective: To investigate the optimal gene transfer protocols of hematopoietic cells mediated by retrovirus. Methods: Murine bone marrow cells were infected by co-culture with murine bone marrow stromal cell line TC-1 or retro-virus packaging cells or retrovirus supernatant. Human mdr-1 and enhanced green fluorescent protein (EGFP) were used as report genes. Results: Stromal cells could greatly increase the gene transfer efficiency when compared with that of supernatant transfection. Transduction efficiency was highest when infected BM cells were co-cultured with virus producer cells. Conculsion: It may be clinically feasible in gene therapy to perform retroviral transduction by co-culture of target cells with stromal cells or cell lines.

Effect of APS on recoverable ability from cryopreservation damage of UCB hematopoietic cells / 中草药

Objective To study the effect of angelica polysaccharides (APS) on recoverable ability from cryopreservation damage of umbilical cord blood (UCB) hematopoietic cells and APS with hematopoietic growth factors (HGFs) on in vitro expansion of UCB mononuclear cells (MNC) after thawing. Methods Thawed UCB MNC were cultured 24 h or 14 d with various concentration of APS (0,50,100,200,and 400 ?g/mL) or with APS and HGFs. The MNC counting assay,typan blue exclusion assay,colorimetric MTT assay for cell proliferation,CFU-Mix colong-forming assay,and flow cytometry for CD34+ cell rate were detected. Results After adding certain concentration APS in thawed UCB cells,the number of MNC,the rate of trypan blue exclusion,the proliferation of MNC,and the colony production of CFU-Mix were significantly enhanced (P

Study on the expression and function of the mdr-1 gene transferred into hematopoietic cells of rabbit bone marrow in vitro / 重庆医科大学学报

Objective:To explore the optimal transfection conditions in which the concentrated virus supernatant transfer the foreign mdr-1 gene into hematopoietic cells of rabbit bone marrow,and test the expression and function of the mdr-1 gene in cells.Methods:An amphotropic virus producer cell line,PA317-HaMDR1/A1 which contains a full-length cDNA of human mdr-1 gene,was screened with colchicine(90ng/ml),and the supernatant was collected and concentrated.The bone marrow cells of the New Zealand rabbits were gathered and the mononuclear cells which contain many hematopoietic cells were enriched.The hematopoietic cells were cocultivated with concentrated virus supernatant and interleukins.The transfection rate was determined by SP immunohistochemical methods;the integration of mdr-1 gene in rabbit mononuclear cells was tested by PCR method ,and the physiological function of mdr-1 gene was tested by DNR extrusion test.Results:After screening by colchicine,the expression of P-glycoprotein(P-gp) of PA317-HaMDR1/A1 was enhanced.The foreign mdr-1 gene was transferred into hematopoietic cells of rabbit bone marrow successfully,and the transduction rate of 2 days,4days and 6 days were 22%,37%,and 39% respectively.However,the state of the 4 days was the best.The transferred mdr-1 gene had its physiological function.Conclusion:The foreign mdr-1 gene can be transferred into hematopoietic cells of rabbit bone marrow successfully by concentrated virus supernatant transfection method,and the transferring gene can be expressed stably and effectively.The study has provided a basis for the further research on chemoprotection experiment of the mdr-1 gene transferred into the bone marrow cells.

Influence of Angelica Sinenisis Injection on Bone Marrow Histology and Ultrastructure in Immune-Induced Aplastic Anemia Mice / 实用儿科临床杂志

Objective To investigate the influence of angelica sinenisis injection on bone marrow histology and ultrastructure in immune-induced aplastic anemia(AA) mice.Methods Female Balb/c mice were divided into normal group,model group,therapy group and contrast group.All mice were killed by cutting neck on the 12 th day, ulnas and femur were taken out. The bone marrow histology section and ultrastructure of mice ulnas were observed. The quantities of hematopoietic cells were counted. The number of femur bone marrow mononucleate cells (MNC) were measured.Results The quantities of hematopoietic cells in model group were lower than those in normal group (P