Basic Information

Common name
B-hB2M/HLA-G MC38
Catalog number
321848
Aliases
MHC-G
Disease
Colon carcinoma
Organism
Mouse
Strain
C57BL/6
Tissue types
Colon
Tissue
Colon
Application
B-hB2M/HLA-G MC38 cells have the capability to establish tumors in vivo and can be used for efficacy studies.
NCBI Gene ID

Description

The mouse B2m gene was replaced by human B2M and HLA-G coding sequence in B-hB2M/HLA-G MC38. Human HLA-G is highly expressed on the surface of B-hB2M/HLA-G MC38 cells.

Targeting strategy

Gene targeting strategy for B-hB2M/HLA-G MC38 cells. The exogenous promoter and human B2M and HLA-G coding sequence was inserted to replace part of murine exon 1 and all of exon 2 and 3. The insertion disrupts the endogenous murine B2m gene, resulting in a non-functional transcript.

Protein Expression Analysis

HLA-G expression analysis in B-hB2M/HLA-G MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hB2M/HLA-G MC38 cells cultures were stained with species-specific anti-HLA-G antibody. Human HLA-G was detected on the surface of B-hB2M/HLA-G MC38 cells but not wild-type MC38 cells. The 2-A08 clone of B-hB2M/HLA-G MC38 cells was used for in vivo experiments.

Tumor growth curve & Body weight changes

Subcutaneous homograft tumor growth of B-hB2M/HLA-G MC38 cells. B-hB2M/HLA-G MC38 cells (5×105) and wild-type MC38 cells (5×105) were subcutaneously implanted into B-Tg(hLILRB2/hLILRB3) mice (female, 7-week-old). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B)  Body weight (Mean± SEM). Volume was expressed in mm3 using the formula: V=0.5 X long diameter X short diameter2. As shown in panel A, B-hB2M/HLA-G MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Protein expression analysis of tumor cells

B-hB2M/HLA-G MC38 cells were subcutaneously transplanted into B-Tg(hLILRB2/hLILRB3) mice (n=3), and on 28 days post inoculation, tumor cells were harvested and assessed for human HLA-G expression by flow cytometry. As shown, human HLA-G was highly expressed on the surface of tumor cells. Therefore, B-hB2M/HLA-G MC38 cells can be used for in vivo efficacy studies of novel HLA-G therapeutics.

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