[Therapeutic cloning and hematopoietic stem cell transplantation].
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Clone Cells↗
Biomedical subjects
Xue-tao Pei
Publications and source records attributed to Xue-tao Pei.
[Sustainment of hepatocyte function with mixed cellular co-encapsulation].
OBJECTIVE: To evaluate the effects of mixed microcapsules of hepatocytes mixed with hepatocytes, transgenic hepatic stellate cell strain (HGF/CFSC), and/or bone marrow derived Thy-1(+) beta(2)M(-) cells (BDTCs) to sustain liver function. METHODS: Three kinds of microcapsules containing hepatocytes, hepatocytes + CFSC/HGFs, or hepatocyte + CFSC/HGF + BDTC were prepared and cultured in conditioned culture fluids. The morphology of the microcapsules and the encapsulated cells were observed by microscopy. The supernatant was collected regularly to detect the secretion of albumin and urea. Forty Wistar rats underwent 90% hepatectomization to establish acute liver failure model. Six hours after the operation the rats were randomly divided into 4 groups to be intraperitoneally injected with one of the 3 kinds of microcapsules containing 3.5 x 10(7) hepatocytes as experimental groups (Groups II, III, and IV) or injected with blank microcapsule as control group. The behaviors of the rats were observed daily. Blood was collected from the eyeball at different time points to detect relevant biochemical indicators. Twenty-one and 42 days after the operation the rats were killed. Abdominal lavage was performed to collect the microcapsules to undergo microscopy. Liver specimens were colleted to undergo pathological examination. RESULTS: Severe liver failure occurred in the rats transplanted with blank microcapsules. Most of the rats in Groups II, III, and IV began to eat within 20 hours after hepatectomization. Nine of the 10 rats in the control group died within 48 hours after hepatectomization. Nine of the 10 rats of Group II survived a long time. All the rats in Groups III and IV survived till the end of experiment. In comparison with the supernatant of Group II, the contents of albumin and urea in the supernatants of Groups III and IV were significantly higher (all P < 0.01). The liver function indicators, ALT, AST, lactic dehydrogenase, and albumin worsened since one day after the operation. Five days after the transplantation of microcapsule, the above indicators showed remarkable improvement, and recovered to normal 7 days after. Twenty-one and 42 days after the transplantation regeneration was seen and edema was reduced in the livers in Groups II, III, and IV. Twenty-one days after the transplantation most of the microcapsules were still free in the peritoneal cavity in Group II. In Group III, most of the microcapsules aggregated around the portal vein, fibrosis at the surface of microcapsule to a certain degree was seen and surviving hepatocytes could be found inside the capsules 21 days after. Forty-two days after, vascularization of microcapsules was seen in Groups III and IV, especially in the latter group. CONCLUSION: Mature hepatocyte, transgenic liver nonparenchymal cells and/or BMSCs co-encapsulate transplantation effectively improve acute liver failure. The microenvironment created by CFSC/HGF and/or BDTC is propitious to the maintenance of capsulated hepatocytes' function and longevity.
Albumins↗
[Ethical problems in human stem cell research and application].
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Humans↗
[Differentiation of rat bone marrow derived Thy-1+ beta2M- cells into hepatocytes].
OBJECTIVE: To investigate the differentiation of bone marrow derived Thy-1(+)beta(2)M(-) cells (BDTC) into mature and functional liver cells and its mechanism. METHODS: BDTC were cocultured with allyl alcohol (AA)-injured hepatocytes and cultured alone in conditional medium containing HGF and bFGF, respectively. BDTC morphologic transformation was observed with phase-contrast and electron-microscopy. Hepatocyte-specific gene expression in cultured BDTC was identified by immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Indocyanine green (ICG) ingestion/excretion and urea, albumin production were carried out to evaluate hepatocyte-related function. RESULTS: Some BDTC derived hepatocyte-like cells with high nuclear to cytoplasmic ratio containing mono- or multi-nuclei and abundant mitochondria, endoplasmic reticulum and glycogenic granules appeared after 7-day culture in both the two culture systems. These cells expressed hepatocyte-specific genes (AFP, OV-6, CK18, etc.), and possessed functions of ICG uptake, albumin production and ammonium metabolism. CONCLUSION: Rat BDTCs could differentiate into mature and functional liver cells in special stimulation systems. Moreover, these differentiations were realized by "transdifferentiation", and might dispense with "cell fusion".
Animals↗
[Hope to the cancer therapy: cancer stem cell].
Cancer stem cells are defined as rare cells in cancer tissues with indefinite potential for self-renewal that drives tumorigenesis. It was first extensively documented for leukaemia and multiple myeloma. It has also been found in solid cancers such as human breast cancer and nervous system tumors. Studies of cancer stem cell biology and mechanisms of tumorigenesis are lending insight into the origins of cancer and will ultimately yield new approaches to fight cancer.
Animals↗
[In vitro differentiation of human bone marrow-derived mesenchymal stem cells into blood vessel endothelial cells].
OBJECTIVE: To study differentiation of human bone marrow-derived mesenchymal stem cells (BDMSC) into blood vessel endothelial cells for ideal cell origin of complex organ tissue engineering vascularization and injured tissue repairing by cell transplantation. METHOD: After different days of induction with vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in 3D fibrin-gels and matrigel, BDMSC and angiogenesis were determined by the utilization of morphological observation, tissue section and CD34, CD31, vascular endothelial growth factor receptor-1 (VEGFR-1, Flt-1), VEGFR-2 (Flk-1), and vWF that were special for blood vessel endothelial cells. RESULT: After 3D-cultured and induced with VEGF and bFGF in vitro in fibrin-gels and matrigel for 3-21 days, BDMSC expressed CD34, CD31, Flt-1, Flk-1, and vWF came into vessel-like configuration. CONCLUSION: VEGF, bFGF as well as Flt-1 and Flk-1, expressed by BDMSC, may form a feasible microenviroment after induction and play an important role during processes of blood vessel endothelial cell differentiation and vessel-like configuration forming of BDMSC. Mesenchymal stem cells may be applied to tissue engineering vascularization and injured tissue repairing by cell transplantation.
Cell Culture Techniques↗
[Autologus bone marrow-derived mesenchymal stem cells intracoronary delivery after acute myocardial infarction in miniature pig].
OBJECTIVE: To evaluate the possibility that using intracoronary delivery of autologus bone marrow-derived mesenchymal stem cells (MSCs) to improve the cardiac function after acute myocardial infarction (AMI) in miniature pig. METHODS: MSCs were cultured in Dulbecco's modified Eagle's medium-F12 (DMEM/F12) medium. AMI model was made by blocking the blood stream of the first diagonal branch in miniature pig, and released the branch after 90 minutes. After 10-14 days, (4-6) x 10(7) culture-expanded autologus 4', 6-diamidino-2-phenylindole (DAPI)-labelled MSCs were transplanted into each host heart's AMI area through intracoronary way. Ultrasonic cardiography (UCG) was performed to observe the left ventricular function at 3 months after transplantation. The cellular transplanted hearts were harvested and investigated by immunohistochemical analysis. RESULTS: Left ventricular function of the MSCs group was improved significantly 3 months later compared with the control group [(54.65 +/- 3.39) vs (43.98 +/- 4.21)%, (P < 0.01)]. Exogenous MSCs survived and site-differentiated into cardiomyocytes in infracted hearts. CONCLUSION: MSCs can play a benificial role to repair damaged heart. Heart function can be improved after MSCs transplantation in porcine myocardial infarction model.
Animals↗
[Migration and differentiation of exogenous rat mesenchymal stem cells engrafted into normal and injured hearts of rats].
OBJECTIVE: To investigate the effects of different microcircumstances on the migration and differentiation of grafted rat mesenchymal stem cells (rMSC) in host myocardium and the feasibility of treatment of myocardial infarction by exogenous adult stem cells. METHODS: rMSC were isolated from the femurs and tibiae of a male Wistar rat and then purified, made into cell suspension, and labeled with DAPI. 35 female Wistar rat were divided randomly into four groups: acute myocardial infarction control group (AMI group, n = 10, the descending anterior branch of left coronary artery was ligated), acute myocardial infarction + rMSC transplantation group (AMI + rMSC group, n = 10, 1 - 3 hours after the ligation DAPI-labeled rMSC were injected into the peri-infarct tissues), normal heart + rMSC transplantation group (normal heart + MSC group, n = 10, DAPI-labeled rMSC were injected into the corresponding myocardium), and mono-nuclear cells transplantation group (AMI + MNCS, n = 5 DAPI-labeled mononuclear cells were injected into he periinfarct tissues). Ten weeks after the implantation, the rats were killed and their hearts were harvested. Immunohistochemistry was used to examine the troponin, GATA-4 and connexin-43. RESULTS: No lymphocyte proliferation and immonologic rejection were seen in the cardiac tissues of the rats implanted with rMSC. DAPI-labeled rMSC with blue nuclei were distributed extensively in the myocardium of the AMI + rMSC group, ovoid in shape and arranged in parallel with the cardiac muscle fibers, and were distributed sporadically like islands in the myocardium of the normal heart + rMSC group, irregular in shape and not arranged in parallel with the cardiac muscle fibers. No blue nucleus was seen in the cardiac tissues of the hearts implanted with DAPI-labeled mononuclear cells. Troponin and GATA4 were positive immunohistochemically in the implanted rMSC with blue nuclei and the host cardiac muscle cells of the AMI group and AMI + rMSC group, however, were negative in the implanted rMSC with blue nuclei and normal cardiac muscle cells of the normal heart + rMSC group. CONCLUSION: Purified rMSC are immunologically tolerable and can be used as donor cells for exogenous cells therapy. Capable of surviving and homing in both in normal and injured hearts, exogenous rMSC migrate and differentiate into cardiac muscle cell-like cells in myocardium with infarction, however, not in normal heart.
Animals↗
[Transplantation of exogenous mesenchymal stem cells for treatment of myocardial infarction in rat].
OBJECTIVE: To investigate the possibility of homing,survival, migration and differentiation of rat mesenchymal stem cells (rMSCs) in host myocardium and the effects of rMSCs transplantation on the function of heart with myocardial infarction; To observe the feasibility of using exogenous adult stem cells for cell therapy. METHODS: 35 female rats were separated randomly into three groups: normal control group (control, n = 10), acute myocardial infarction control group (AMI, n = 10) and myocardial infarction plus cell transplantation group (AMI + Cell, n = 15). The infarcted hearts were made by occlusion of left coronary artery. Male Wistar rats MSCs were isolated and purified. Then the cells were implanted into the infarcted hearts of female Wistar rats. Heart functions were measured 10 weeks after implantation. The hearts were harvested 10 weeks after implantation for immunohischemistry. RESULTS: (1) Purified rMSCs survived and homed in exogenous host hearts; (2) DAPI-labelled rMSCs with oval nucleus were widely distributed and stained positively of cardiac specific proteins. The arrangements of the donated cells paralleled with host myocardium fibers in infarcted host hearts. Transplantation of rMSCs was associated with a significant decrease of end left ventricular volumes, increase of left ventricular end-systolic pressure and increase of ratio of left ventricular pressure rise and left ventricular pressure decay (+/- dp/dt) (P < 0.05) as compared with the control hearts. The numbers of blood vessels were increased at the boundary of infarction site and the sizes of infarction were decreased significantly (P < 0.05). CONCLUSIONS: The purified rMSCs can survive in exogenous host hearts without addition of any immunosuppressant. These exogenous cells take part in myocardium regeneration and have beneficial effects on heart function.
Animals↗
[Observation of bone marrow mesenchymal stem cells after subretinally transplanted into laser-injured rat retina].
OBJECTIVE: Bone marrow mesenchymal stem cells (MSCs) develop into hematopoietic and mesenchymal lineages but have not been known to participate in production of retina. Eye was known as an immunologically privileged organ. Because of its special structure, it's difficult for blood cells to enter retina. This article is to trace bone marrow mesenchymal stem cell (MSCs) after subretinally transplanted into Nd: YAG laser-injured rat retinal without in vitro differentiation induction. METHOD: 4', 6-diamidino-2-phenylindole (DAPI)-labeled MSCs were used to trace the change of MSCs after transplantation on day 10, 20, 35 and 50. The sequenced sections were used for Immunohistochemistry identification for neuronal nuclei (NeuN), neuron specific enolase (NSE), glial fibrillary acidic protein (GFAP) and pancytokeratin (CK). Electroretinogram (ERG) b waves were recorded for each eye of the laser injured group, the laser injured transplanted group and the laser injured with saline injection control group before, right after or at every end of 1 to 7 weeks. RESULTS: On day 10, the DAPI positive cells were mainly crowded around the transplanted site. On day 20 the positive area enlarged and scattered into RPE layer, photoreceptor layer, bipolar layer and ganglion cell layer. Then the positive area enlarged more widely on day 35 and more cells could be found migrate to the lesion site. The positive area didn't enlarge much on day 50 than on day 35. No formation of rosettes was found during the observation. But the expression of NeuN, NSE, GFAP and CK was not uniform as the normal retina. Cell proliferation was still found. But HE staining showed that the lesion in the transplanted group was better than that of the control group. Correspondingly ERG b-wave value at week 5 was higher than the control group. CONCLUSION: From these cells, a proportion of the cells regenerated from bone marrow can be differentiated into retina in vivo. Although the MSCs-derived cells could not precisely express neuro-like proteins, they could help recover lesion and ERG b-wave value.
Animals↗
[Immune response of dendritic cells acquiring antigens from apoptotic cholangiocarcinoma cells induced by mitomycin].
OBJECTIVE: To investigate the anti-tumor immune response of dendritic cells (DCs) acquiring antigens from apoptotic cholangiocarcinoma cells and their therapeutic effects on cholangiocarcinoma cells. METHODS: DCs from human peripheral blood monocytes which acquired antigen capturing and processing capacity, characteristics of maturation, were established in vitro using granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4). Then cholangiocarcinoma cells were induced to apoptosis with mitomycin. The three groups included (1) coculture of DCs, apoptotic cancer cells and T cells, (2) coculture of DCs, necrotic cancer cells and T cells, (3) coculture of DCs, cultured cancer cells and T cells. After 7 days, DCs and T cells were riched separately to perform anti-tumor cells test and immune response test. RESULTS: these cells had typical dendritic cell morphology, expressed high levels of CD1a and B7, acquired antigen from apoptotic cells caused by mitomycin and could stimulate T cells to inhibit, even kill cholangiocarcinoma cells. CONCLUSIONS: The DCs from peripheral blood monocytes induced by GM-CSF and IL-4 can efficiently present antigen derived from apoptotic cells caused by mitomycin, and stimulate T cells activity obviously. It maybe become an effective therapy for tumor.
Antigens, Neoplasm↗
[Identification of differentially expressed genes in Lin-CD34- and Lin-CD34+ cells].
OBJECTIVE: To identify genes that differentially expressed in Lin(-)CD(34)(-) and Lin(-)CD(34)(+) cells. METHODS: With Lin(-)CD(34)(-) cells as tester and Lin(-)CD(34)(+) cells as driver, cDNA subtractive library for Lin(-)CD(34)(-) cells was constructed using suppression subtractive hybridization technique. Part of clones in the library were sequenced and the homologue analysis was conducted against the DNA database in GenBank. RESULTS: 593 clones containing an average of 300 - 500 bp insert were identified. Of them, 53 randomly selected ESTs were sequenced. Homologue analysis revealed that 37 ESTs represented 10 known genes, and the other 16 ESTs represented 4 novel sequences. CONCLUSION: Part of specifically expressed genes in Lin(-)CD(34)(-) cells were identified, which maybe related to Lin(-)CD(34)(-) cells' specific characteristics.
Antigens, CD34↗
[Effect of Tpo and/or IL-11 gene modified stromal cells on the expansion of CD34+ CD38- hematopoietic primitive progenitor cells].
OBJECTIVE: To investigate the effects of Tpo and/or IL-11 gene modified stromal cells on the expansion of CD(34)(+) hematopoietic stem/progenitor cells in cord blood. METHODS: Retroviral vectors containing Tpo or IL-11 gene were constructed and used to transfect the stromal cell line HFCL. Tpo and/or IL-11 mRNA was assayed by Northern blot. Non-modified stromal cells were used, CD(34)(+) hematopoietic stem/progenitor cells from cord blood were expanded on gene-modified stromal cells for 7 days. The phenotype of CD(34)(+)CD(38)(-) primitive progenitors was detected by flow cytometry. RESULTS: HFCL expressed Tpo and/or IL-11 mRNA after transfected by the retroviral vectors. The percentages of CD(34)(+)CD(38)(-) primitive progenitors in the cultures of Tpo, IL-11 and Tpo + IL-11 modified HFCL were (1.8 +/- 0.24)%, (1.62 +/- 0.23)%, and (2.45 +/- 0.28)%, respectively, which were higher than that in the control [(0.8 +/- 0.23)%]. CONCLUSION: The stromal cells modified by Tpo and/or IL-11 gene were able to enhance ex vivo expansion of CD(34)(+) and CD(34)(+)CD(38)(-) hematopoietic stem/progenitor cells from cord blood.
ADP-ribosyl Cyclase↗