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B-NDG MGMT3 mice
Strain Name 

NOD.CB17-Prkdcscid Il2rgtm1Bcgen Il3tm1(IL3)Bcgen Csf2tm1(CSF2)Bcgen 

Csf1tm1(CSF1)Bcgen Thpotm1(THPO)Bcgen/Bcgen

Common Name 

B-NDG MGMT3 mice

Background B-NDG mice Catalog number 111882
Aliases 

IL3: IL-3, MCGF, MULTI-CSF
Csf2: CSF, GMCSF
Csf1: CSF-1, MCSF
THPO: MGDF, MKCSF, ML, MPLLG, THCYT1, TPO

Protein expression analysis

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Strain specific GM-CSF, CSF1 and THPO expression analysis in wild-type B-NDG mice and homozygous B-NDG MGMT3 mice by ELISA. Serum was collected from the two mice stimulated with LPS in vivo and analyzed by ELISA (n=3). Mouse GM-CSF, CSF1 and THPO were only detectable in B-NDG mice (+/+) but not in B-NDG MGMT3 mice (H/H). Human GM-CSF, CSF1 and THPO were only detectable in B-NDG MGMT3 mice. As IL3 is mainly expressed in activated T cells and there is no mature T cells in B-NDG background mice, mouse or human IL3 was not detectable in both of the two mice.


Analysis of leukocyte subpopulation in spleen


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Analysis of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from male B-NDG and B-NDG MGMT3 mice (n=3, 7-8-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percentages of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes and macrophages in homozygous B-NDG MGMT3 mice were similar to those in the B-NDG mice, demonstrating that IL3, GM-CSF, CSF1 and THPO humanized does not change the overall development, differentiation or distribution of these cell types in spleen. Values are expressed as mean ± SEM.

Analysis of leukocyte subpopulation in blood

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Analysis of leukocyte subpopulations in blood by flow cytometry. Blood was isolated from male B-NDG and B-NDG MGMT3 mice (n=3, 7-8-week-old). Flow cytometry analysis of the blood cells was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percentages of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes and macrophages in homozygous B-NDG MGMT3 mice were similar to those in the B-NDG mice, demonstrating that IL3, GM-CSF, CSF1 and THPO humanized does not change the overall development, differentiation or distribution of these cell types in blood. Values are expressed as mean ± SEM.

Analysis of leukocyte subpopulation in bone marrow

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Analysis of leukocyte subpopulations in bone marrow by flow cytometry. Bone marrow was isolated from male B-NDG and B-NDG MGMT3 mice (n=3, 7-8-week-old). Flow cytometry analysis of the cells from bone marrow was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, dendritic cells, neutrophils, monocytes and macrophages in homozygous B-NDG MGMT3 mice were similar to those in the B-NDG mice, demonstrating that IL3, GM-CSF, CSF1 and THPO humanized does not change the overall development, differentiation or distribution of these cell types in bone marrow. Values are expressed as mean ± SEM.




Human CD34+ HSCs engraftment for human immune system reconstitution  


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Human CD34+ HSCs (3E4) were intravenous (temporal vein) engrafted into wild-type B-NDG mice and homozygous B-NDG MGMT3 mice (both sex, 24-72 hr after birth, n=15). B-NDG mice were treated with 1.0 Gy-irradiation. B-NDG MGMT3 mice were not irradiated. A. Survival rates of the mice were analyzed with Kaplan Meier survival curves. B. Body weight. Results showed that the survival rate of B-NDG MGMT3 mice was similar to that of B-NDG mice until 18 weeks after human CD34+ HSCs engraftment and then decreased to 42.85% at 24 weeks post engraftment. But the body weight of B-NDG MGMT3 mice was significantly higher than that of B-NDG mice and increased steadily during the whole reconstitution. Values are expressed as mean ± SEM. HSCs: hematopoietic stem cells.


Human CD34+ HSCs engraftment for human immune system reconstitution  

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Human CD34+ HSCs (3E4) were intravenous (temporal vein) engrafted into wild-type B-NDG mice and homozygous B-NDG MGMT3 mice (both sex, 24-72 hr after birth, n=15). B-NDG mice were treated with 1.0 Gy-irradiation. B-NDG MGMT3 mice were not irradiated. Peripheral blood lymphocytes from the two mice after engraftment with human CD34+ HSCs were analyzed with flow cytometry. Results showed that the proportion of CD45+ cells in B-NDG MGMT3 mice reached 25% starting from 12 weeks after engraftment and continued to rise, significantly higher than that in B-NDG mice. The proportions of monocytes, MDSCs, DCs and Tregs in B-NDG MGMT3 mice were higher than that in B-NDG mice. Values are expressed as mean ± SEM. 

Human CD34+ HSC engraftment for human immune system reconstitution  

from clipboard


Human CD34+ HSCs (3E4) were intravenous (temporal vein) engrafted into wild-type B-NDG mice and homozygous B-NDG MGMT3 mice (both sex, 24-72 hr after birth, n=15). B-NDG mice were treated with 1.0 Gy-irradiation. B-NDG MGMT3 mice were not irradiated. Peripheral blood lymphocytes from the two mice after engraftment with human CD34+ HSCs were analyzed with flow cytometry. Results showed that the proportion of CD45+ cells in B-NDG MGMT3 mice reached 25% starting from 12 weeks after engraftment and continued to rise, significantly higher than that in B-NDG mice. The proportions of monocytes, MDSCs, DCs and Tregs in B-NDG MGMT3 mice were higher than that in B-NDG mice. Values are expressed as mean ± SEM. 

Human CD34+ HSC engraftment for human immune system reconstitution  


from clipboard


Human CD34+ HSCs (3E4) were intravenous (temporal vein) engrafted into wild-type B-NDG mice and homozygous B-NDG MGMT3 mice (both sex, 24-72 hr after birth, n=15). B-NDG mice were treated with 1.0 Gy-irradiation. B-NDG MGMT3 mice were not irradiated. Peripheral blood lymphocytes from the two mice after engraftment with human CD34+ HSCs were analyzed with flow cytometry. Results showed that the cell numbers of all the cells analyzed from 12 weeks after engraftment in B-NDG MGMT3 mice were higher than that in B-NDG mice. Values are expressed as mean ± SEM. Values are expressed as mean ± SEM. 

Human CD34+ HSC engraftment for human immune system reconstitution  

from clipboard

Human CD34+ HSCs (3E4) were intravenous (temporal vein) engrafted into wild-type B-NDG mice and homozygous B-NDG MGMT3 mice (both sex, 24-72 hr after birth, n=15). B-NDG mice were treated with 1.0 Gy-irradiation. B-NDG MGMT3 mice were not irradiated. Peripheral blood lymphocytes from the two mice after engraftment with human CD34+ HSCs were analyzed with flow cytometry. Results showed that the cell numbers of all the cells analyzed from 12 weeks after engraftment in B-NDG MGMT3 mice were higher than that in B-NDG mice. Values are expressed as mean ± SEM. Values are expressed as mean ± SEM.