Immortalized Human Endometriotic Cell Line (12Z)
Description
| Description | Immortalized Human Endometriotic Cell Line (12Z) was generated from cells obtained from a 37-year-old female undergoing laparoscopy and immortalized by plasmid-mediated expression of SV40 T-antigen (late region deleted, bp 1782–2533) delivered via electroporation. This cell line retains expression of markers characteristic of endometriotic lesions observed in vivo. Immortalized Human Endometriotic Cell Line (12Z) may be used to characterize and identify genes and/or markers related to endometriosis. |
| Organism | Human (H. sapiens) |
| Tissue | Endometrium |
| Donor History | Female, 37 |
| Growth Properties | Adherent, epitelial-like |
| Storage Condition | Vapor phase of liquid nitrogen, or below -130°C. |
| Product Format | Frozen |
| Intended Use | This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. |
| BioSafety | II |
| Certificate of Analysis | For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com. |
| Growth Conditions | For optimal cell culture, we recommend using PriCoat™ T25 Flasks   (G299)   or coating your preferred vessels with Applied Cell Extracellular Matrix   (G422).    PriGrow III (TM003) + 10% FBS (Regular*) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂
*Do not heat-inactivate Batch testing of FBS is recommended as different batches may impact proliferation and marker expression. |
| Unpacking and Storage Instructions | 1. Visually examine the packaging containers for signs of leakage or breakage.
2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130°C, preferably in liquid nitrogen vapor phase storage, until ready for use. To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability. |
| Thawing Protocol | 1. Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.
2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions. 3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes. 4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask. 5. Incubate the cells at the recommended conditions. |
| Subculture Protocol | Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent.
1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment. 3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired. 6. Incubate the cells at the recommended conditions. |
| Cryopreservation | We recommend using serum-free CryoGuardâ„¢ Â Freezing Media (TM078)Â or, if serum is preferred, Cryopreservation Medium (TM024). |
| 3D Culture Conditions |
ProcedureÂ
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| Seeding Density (cells/cm2) | 15,000 – 20,000 |
| Split Ratio | 1:6 or 1:8 |
| Population Doubling Time (h) | 55 – 65 |
| Immortalization Method | Immortalized by plasmid-mediated SV40 T-antigen expression (late region deleted, bp 1782–2533) delivered via electroporation. |
| Expression | SV40T, cytokeratin, vimentin, N-cadherin, estrogen receptor α and β, progesterone receptor, aromatase P450 |
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