CERV-186 Cells
General information
Description | The CERV-186 cell line, derived in-vitro from the xenotransplant cervix carcinoma MRI-H-186, represents a model of invasive, large cell, non-keratinizing squamous cell carcinoma. This establishment and adaptation to in vivo transplantation were facilitated by Dr. Bodgen at the Mason Research Institute. MRI-H186 cells are characterized by their genomic composition, harboring approximately 26 integrated copies of both full-length and truncated HPV16 genomes, which is reflected in their transcriptomic profile. These cells display a pronounced expression of both full-length and truncated early HPV16 transcripts, with a particularly high expression level of E5 full-length (fl) RNA. This expression profile markedly differs from that observed in the CaSki and MRI-H196 cell lines. Moreover, the transcriptional activity in MRI-H186 cells, in terms of the expression of various other transcripts, aligns closely with that seen in the HPK-IA and C3 cell lines, suggesting a shared pattern of transcriptional behavior. The presence of both full-length and truncated HPV16 genomic integrations in MRI-H186 cells underlies their vigorous expression of early viral transcripts, underscored by the significant expression of E5 fl RNA. This indicates the transcription of full-length early RNAs, culminating at the early polyadenylation signal, and highlights the unique transcriptional dynamics within the MRI-H186 cell line. |
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Organism | Human |
Tissue | Cervix |
Disease | Squamous cell carcinoma |
Synonyms | Cerv-186, MRI-H-186, MRI-H186 |
Characteristics
Age | 42 years |
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Gender | Female |
Ethnicity | African |
Morphology | Epithelial-like |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | CERV-186 (Cytion catalog number 300290) |
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Biosafety level | 2 |
Expression / Mutation
Tumorigenic | Yes, in nude mice |
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Viruses | HPV-16 positive |
Products | Cytokeratine 8, 18, Vimentin, Desmoplakin |
Handling
Culture Medium | DMEM:Ham's F12, w: 3.1 g/L Glucose, w: 1.6 mM L-Glutamine, w: 15 mM HEPES, w: 1.0 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | A ratio of 1:2 to 1:6 is recommended |
Seeding density | 2 x 10^4 cells/cm^2 will result in a confluent monolayer within 7 days |
Fluid renewal | 2 to 3 times per week |
Freezing recovery | After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile |
Amelogenin: x,x
CSF1PO: 9,11
D13S317: 12
D16S539: 13
D5S818: 11,12
D7S820: 8,12
TH01: 6
TPOX: 8,11
vWA: 14,17
D3S1358: 15,18
D21S11: 29,30
D18S51: 16
Penta E: 5,7
Penta D: 10,12
D8S1179: 14
FGA: 19,20
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HLA alleles |
A*: 30:01:01
B*: 13:02:01
C*: 06:02:01
DRB1*: 07:01:01
DQA1*: 02:01:01
DQB1*: 02:02:01
DPB1*: 03:01:01
E: 01:01:01
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