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Objective To set up a favorable animal model for the drug treatment research of endometriosis by establishing the animal model of endometriosis in SCID and nude mice so as to compare the influences on implantation of human endometrial tissue derived from the eutopic and ectopic sources. Methods Eutopic and ectopic endometrium were transplanted to the lower abdominal parts subcutaneously of 30 sexually matured BALB/c-nu/nu nude mice and SCID mice respectively. The ectopic lesion sizes were under the regular observation before they were removed 6 weeks after the operation for pathological examinations. Results Nude mice and SCID mice were able to be used to establish a successful animal models of endometriosis. The success rate of SCID mice was higher than that of nude mice. The success rate of the eutopic endometrium group was significantly higher than that of the ectopic endometrium group. Nude and SCID mice endometriosis implantation models were successfully established. The modeling success rate of SCID mice is higher than that of the nude mice.The success rate of transplantation was higher in the ectopic endometrium than in the eutopic endometrium.Conclusion The SCID mice endometriosis endometriosis model provides a favorable animal model of endometriosis.
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Objective To observe the changes in estrous cycle and vaginal smears in ovarectomized NOD/SCID mice.Methods To continuously observe the estrous cycle time by vaginal smears of NOD/SCID mice in consecutive nine days, twice daily.After ovariectomy, the changes of estrous cycle were observed by vaginal smears for 7 days.Results The estrous cycle in NOD/SCID mice was 4-6 days.Regular estrous mice accounted for 80%.There was no significant correlation between vaginal opening and estrous cycle status.After ovariectomy, the vaginal smears showed characteristics of metestrus or diestrus.Conclusions Vaginal smear cytology can be used to determine the estrous cycle and characteris-tics of NOD/SCID female mice.The ovariectomized operation of NOD/SCID female mice is effective.
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Objective To investigate the potential of chronic myeloid leukemia ( CML) cell line KCL22 in indu-cing leukemia in NOD-SCID mice for setting up a basis for constructing a CML mouse transplantation tumor model. Methods 2 ×107 KCL22 cells in logarithmic growth phase were injected via the tail vein into experimental NOD-SCID mice whereas PBS was injected to the mice of control group.General condition of the mice of both groups was observed.Wright staining was used to observe the changes of blood and bone marrow smears.PCR was conducted to detect the transcription level of BCR-ABL, and histology with HE staining was used to evaluate the tumor cell invasion in the liver and spleen. Results Four weeks after the injection of KCL22 cells, the mice in experimental group showed physical signs of decreased reactivity, depression, swollen hindlimb muscles and petechia on the hindlimb femur.Peripheral white blood cells ( WBC) began to increase after 5 weeks, with a significantly increased quantity compared with the control group (P90 days) (P<0.05).Conclusions A NOD-SCID mouse model of CML transplantation tumor is successfully established with leukemia KCL22 cells.
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Objective To measure the organ weights , physiological and biochemical parameters and immune cells of NOD/SCID mice.Methods Mice at five and ten weeks of age were selected .The organ weights, blood physiological and biochemical parameters were observed .The percentages of CD 3+, CD4+, CD8+, CD19+, B220+, NK1.1+, and CD11b+were checked by FCM in NOD/SCID mice at six week of age in terms of its T , B lymphocyte function and NK cell activity.Results Among the same sex group of NOD/SCID mice, the weights of kidney, liver, heart and lung, and the blood physiological indexes of RBC , HGB, HCT, MCV, MCH, RDW, and the blood biochemical indexes of TP , ALB, ALP, CHO, TBIL show significant difference between 5 and 10 weeks.At the same age, HCT, GLOB, A/G, CHO, TG, TBIL and the UN are significantly different between male and female NOD /SCID mice.NOD/SCID mice lack T cells (0.37 ±0.26)%、CD4+T cells (0.35 ±0.13)%、CD8 +T cells (0.47 ±0.10)%、CD19+B cells (0.13 ± 0.05)%、B220+B cells (1.20 ±0.44)%.The percentage of NK cells is (6.90 ±0.82)%, and the percentage of granulocytes is (47.88 ±15.54)%.Conclusions The study indicates that NOD/SCID mice show the deficiency of T , B and NK cells function .The organ weights , blood physiological and biochemical parameters are affected by age and gender . The studied parameters of NOD/SCID mice are similar with the same strain in other countries .
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Purpose To study the malignant transformation after treating rat oval cell line ( WB-F344 ) with chemical carcinogen N-methyl-N′-nitro-N-nitrosoguanidine ( MNNG) . Methods WB-F344 cells were cultured with MNNG for severe times. The biological characteristics of induced cells were detected through the following methods:to check proliferation activity by flow cytometry analysis, to examine malignant transformation degree of induced cells by soft agar assay and tumor formation in NOD/SCID mice, and to investi-gate the transcriptional and protein levels of hepatocellular carcinoma marker GGT, GST-P by real time-PCR. Results Oval cells in-duced by MNNG showed changes in biological characteristics and malignant molecular markers. Conclusion Hepatic oval cells model is successfully established, which can be confirmed by tumor formation in NOD/SCID mice.
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Objective To compare the characteristics of hMPV infection in BALB/c and SCID mice.Methods BALB/c and SCID mice were infected intranasally with GFP-rhMPV,and sacrificed on day 3,5,7,9 and 14 post inoculation.Heart,liver,spleen,lungs,kidneys and brain of the animals were used for viral isolation,titration,pulmonary histopathology and detection of GFP-hMPV mRNA expression by RT-PCR and real-time PCR.Results Live viruses were successfully isolated from the lungs of infected mice.Viral titers peaked on the 5th day post inoculation.Viruses remained to be detectable on the 14th day post inoculation in SCID mice,but not in BALB/c mice,whereas genomic RNA of GFP-rhMPV was detectable by PCR targeting F gene in infected BALB/c mice.Live viruses were not able to be isolated from heart,liver,spleen,kidney and brain,neither was genomic RNA of hMPV able to be detected on the 5 th day post inoculation by RT-PCR and real-time PCR.Histopathology of lungs was characterized by interstitial pneumonia on 5 days post inoculation.Lung pathology score of BALB/c mice group was slightly lower than SCID,and the difference was not statistically significant.Conclusion GFP-rhMPV can only replicate in the immunocompetent and immunodeficient mouse lungs,but not in other organs.As compared to that in BALB/c mice,the viral replication appears to be more efficiently and for longer time in SCID mouse lungs probably due to the absence of host cellular and humaral immunity,but this does not necessarily result in more severe pathological lesion.
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Objective To establish Ad-hLIF transgenic feeder cells for the expansion of umbilical cord blood CD34+ HSPC in vitro and study the SCID mice model of hematopoietic stem/progenitor cell (HSPC) transplantation. Methods The expression of objective gene in Ad-hLIF transgenic feeder cells was detected by RT-PCR and ELISA. The purity of umbilical cord blood CD34+ HSPC separated by magnetic-activated cell sorting(MACS) was detected by the flow cytometry. After expanded with various combinant of cytokines and transgenic feeder layer cells for 28 d, the quantity of mono-nuclear cell (MNC) and CD34+ cells rate was detected in different time. MNC after expansion stained by CFDA SE was injected to the sublethally irradiated SCID mice. Humanize gene Alu was detected by RT-PCR and fluorescence microscope. Results The green fluorescence was observed in the transgenic cells infected with 50MOI( multiplicity of infection) Ad-hLIF, and the objective gene was confirmed by RT-PCR and ELISA. The purity of umbilical cord blood CD34+ HSPC separated by MACS could reach 95.60% ±2.58%, Ad-hLIF transgenic feeder cells and various cytokines system increased MNC by 356.95±0.87 fold, and maximal expansion of CD34+ cells was observed during 0-14 d of culture, then down-expansion gradually. Four weeks after transplanted in SCID mice, fluorescently-labeled humanize cells still can be observed. The existence of the humanized gene Alu was confirmed by RT-PCR. Conclusion Ad-hLIF transgenic feeder cells can effectively proliferate umbilical cord blood CD34+ HSPC in vitro and delay it differentiate, what's more, it has high transplant efficacy and haematogenesis activity.
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Objective:To investigate the feasibility of establishing a human renal cell carcinoma model in human peripheral blood lymphocyte-engrafted to severe combined immunodeficient(hu-PBL-SCID)mice.Methods:The biological and immunological features of mice were evaluated after intra-peritoneal injection of human peripheral blood lymphocytes(PBL)and subcutaneous implantation of human renal cell carcinoma cells(RCCCs).Results:(1)Subcutaneous tumors developed in all the mice given human PBL and RCCCs.The latency period was significantly prolonged,and the tumor size was markedly depressed,as compared with the mice given RCCCs(P
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Objective To study the roles of dengue (DEN) virus specific T cells in the pathogenesis of DEN virus infection. Methods 2D42 cells, DEN virus specific CD8 + cell clones, were employed to investigate their in vivo function in DEN virus infection using an animal model. HepG2 was implanted into mice with severe combined immunodeficiency disease (HepG2-SCID) for the establishment of HepG2-SCID model. The animals were divided into 3 groups: Group A: HepG2-SCID mice were inoculated with 2D42 cells and then infected with DEN virus type 2 (DEN2) intraperitoneally; Group B: HepG2-SCID mice were inoculated with normal mouse thymuscytes (NMT) and then intraperitoneally infected with DEN2; Group C: HepG2-SCID mice were intraperitoneally infected with DEN2 alone. The mortality, viremia, and frequency of histopathological changes in the major organs of mice in the three groups were observed after infection. Results After inoculation of 2D42 cells, 80% infected mice showed severe clinical signs and died at the average 12.8 d after infection. The others only had transient manifestations, and then recovered from the disease and survived for more than 3 months. In contrast, after inoculation of NMT and /or DEN2 alone, 100% mortality rate was noted in these two groups. High viremia and frequency of histopathological changes in the major organs were observed in the mice in groups A and B. Conclusion Our data support both protective and pathogenic roles for DEN-specific CD8 + T cells in DEN virus infection.
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Attempts were made to isolate and identify Korean bovine Babesia parasite. Blood samples were collected from Holstein cows in Korea, and Babesia parasites were propagated in SCID mice with circulating bovine red blood cells for isolation. The isolate was then antigenically and genotypically compared with several Japanese isolates. The Korean parasite was found to be nearly identical to the Oshima strain isolated from Japanese cattle, which was recently designated as Babesia ovata oshimensis n. var. Haemaphysalis longicornis was the most probable tick species that transmited the parasite.
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Animals , Mice , Arthropod Vectors/parasitology , Babesia bovis/genetics , Babesiosis/parasitology , Base Sequence , Cattle/parasitology , Cattle Diseases/parasitology , DNA, Protozoan/genetics , DNA, Ribosomal/genetics , Erythrocytes/parasitology , Korea , Mice, SCID , Molecular Sequence Data , Phylogeny , Ticks/parasitologyABSTRACT
Objective:To study the effect of Cyclosporine A(CSA) on inhibiting graft versus host reaction(GVHR) occured in hu PBL/SCID chimeras and to stably establish EBV induced lymphoma models.Methods:Human peripheral blood lymphocyts were isolated and were inoculated intraperitoneally into SCID mice.Mice were infected with EBV and injected intraperitoneally with CSA.Human sIL 2R in the serum of hu PBL/SCID chimeras were analyzed by ELISA.Results:No mouse was dead in CSA group,whereas 15 mice of the other three groups died of GVHR.The medium life span of no CSA administration mice was 17 days,and motalities were 55.56%(5/9),30.43%(7/23),42.86%(3/7)respectively.The difference was statistically significant between CSA group and the other groups.The levels of human sIL 2R were stable in CSA group while increased gradually in experimental infertion the groups without CSA.Difference was significant at day 15 and day 22 between the EBV infection group without CSA and with CSA administration.Of 38 survival SCID mice,24 mice developed tumors in their body cavities.Conclusion:CSA can strikingly inhibit GVHR that may occur in hu PBL/SCID mice,that could help practical to stably establish the lymphoma models.
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AIM: To observe the effects of valproic acid(VPA) and HA14-1 on Bcl-2 overexpressed leukemia cells in vitro and in vivo.METHODS:(1) Cells were divided into control group,HA14-1 group,VPA group and HA14-1+VPA group.The apoptotic rate,the mean fluorescent index(MFI) of Bcl-2,the levels of caspase 3,8 and 9 were detected with FCM.(2) 24 h after transplantation with BALL-1,the NOD/SCID mice were divided into 4 groups as(1),then the survival time was compared.The expression of CD19 in the peripheral blood cells,bone marrows,livers,spleens and lungs of each group was detected.RESULTS:(1) The apoptotic rate in HA14-1+VPA group was(76.5?6.9)%,this was significantly elevated compared to the data in other groups(P
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AIM: Humanized-NOD/SCID(hu-NOD/SCID) mouse model was established and the level of immune reconstitution was assessed in this model. METHODS: Mononuclear cells (MNC) and CD34+ cells were isolated or sorted from cord blood(CB). Human CD45, CD19, CD3 markers on cells from NOD/SCID murine peripheral blood(PB), bone marrow(BM), thymus were detected by FCM from 4 to 10 weeks after hematopoietic stem cell transplantation. After 10 weeks, the gene expressions of the human ?2M and RAG2 were detected by RT-PCR in PB or bone marrow of mice model. RESULTS: Human CD45, CD19, CD3 cells populations in PB and BM were found by flow cytometry in mice model transplanted with CD34+ cells or CB MNC from 4 to 10 weeks. The highest positivity of human lymphocytes was at 8 week after transplantation. The levels of human cell engraftment in mice transplanted with CD34+ cells were higher than those in mice transplanted with CB MNC. The mRNA of human ?2M and RAG2 were found by RT-PCR in BM.CONCLUSION: The higher level of human lymphocyte engraftment is established in NOD/SCID mouse model transplanted with CD34+ compared with CB MNC. The maturation of T lymphocytes could be happened in bone marrow of mice model.
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Objective:To detect the engraftment capability and hematopoietic potential of the expanded cells,establish an optimized project of ex vivo expansion and the SCID mice model of UCB transplantation.Methods:Human cord blood CD34 + cells were clutured in strom free liquid culture system,the sublethally irradiated SCID mice were injected with human expanded cord blood cells by tail vein.Four weeks after inocuration,BM cells of the survived animals are obtained from both femurs and tibias and assessed for the presence of human cells by immunofluorescence staining and PCR.Results:The cells and CFUs of group FL+TPO+SCF+IL 6 were increased significantly,and maintained some proportional CD34 + cells.The CFU of group SCF+IL 6+IL 3+GM CSF+EPO had reduced in week 2,CFC and CD34 + cells and Alu gene were detected in SCID mice BM.Conclusion:The cytokine combination with FL+TPO+SCF+IL 6 can effectively expand UCB CD34 + cells,and the expanded cells can engraft the BM of sublethally irradiated SCID mice and reconstitute hematopoiesis.
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Objective Choose an easy method to screen the “leaky" phenotype SCID mice before experimental use. Methods IgG in SCID mice serum was assayed by three kinds of ELISA(sandwich ELISA, sandwich indirect ELISA and Avidin\|biotin sandwich ELISA). Results Avidin\|biotin sandwich ELISA is the most sensitive method of the three, and 41 SCID mice of 6～8 weeks old were assayed with this method. The average IgG of 40 SCID mice was 0 24?0 16mg/L, only one was 2^3mg/L. The “Leaky" probability was 2^4%.Conclusion\ The Avidin\|Biotin sandwich ELISA is an easy, sensitive and reliable method to screen“leaky" SCID mice.\;[