KC-3790-DW

MC38-E6-E7 Cell Line

57645

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Background of MC38-E6-E7 Cell Line

E6 and E7, two major oncogene.Since E6 and E7 are the biomarkers of a cervical cancer cell and are the ones driving the cancer progression, therapeutic approaches targeting E6 and E7 have been proved to be highly efficient in terms of focused removal of abnormally propagating malignant cells. Human papillomavirus (HPV) is a class of envelope-free double-stranded DNA virus. HPV infection has been strongly associated with the development of many malignancies, such as cervical, anal and oral cancers. The viral oncoproteins E6 and E7 perform central roles on HPV-induced carcinogenic processes,HPV-E6 and E7 oncoproteins, considered as biomarkers usable in managing screen-positive women.

Specifications

Catalog Number	KC-3790-DW
Cell Line Name	MC38-E6-E7 Cell Line
Clone Number	4#
Host Cell Line	MC38
Description	Stable MC38 cell line expressing exogenous E6 and E7 gene
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 5 μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

MC38-E6-E7 Cell Line was generated using a lentiviral vector expressing the E6 and E7 sequence.

Characterization

Figure 1: Characterization of E6 and E7 overexpression in the MC38-E6-E7 stable clone using FACS.

Figure 2: Characterization of E6 and E7 overexpression in the MC38-E6-E7 stable clone using PCR.

Figure 3:Validation of in vivo tumorigenicity of MC38 cells stably expressing E6 and E7 via subcutaneous implantation in C57BL/6J mice, with tumor growth monitored by measuring volume (V=0.5×length×width²) and body weight.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS and 5 μg/mL Puromycin)in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage.

References

Pal A, Kundu R. Human Papillomavirus E6 and E7: The Cervical Cancer Hallmarks and Targets for Therapy. Front Microbiol. 2020 Jan 21;10:3116.
Peng Q, Wang L, Zuo L, Gao S, Jiang X, Han Y, Lin J, Peng M, Wu N, Tang Y, Tian H, Zhou Y, Liao Q. HPV E6/E7: insights into their regulatory role and mechanism in signaling pathways in HPV-associated tumor. Cancer Gene Ther. 2024 Jan;31(1):9-17.
Downham L, Jaafar I, Rol ML, Nyawira Nyaga V, Valls J, Baena A, Zhang L, Gunter MJ, Arbyn M, Almonte M. Accuracy of HPV E6/E7 oncoprotein tests to detect high-grade cervical lesions: a systematic literature review and meta-analysis. Br J Cancer. 2024 Mar;130(4):517-525.