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Cell line: | HCEC-B4G12 |
DSMZ no.: | ACC 647 |
Species: | human (Homo sapiens) |
Cell type: | corneal endothelium |
Origin: | HCEC-B4G12 is a clonal subpopulation started in 2005 from the parental cell line HCEC-12 (ACC 646) established from normal cells of the posterior epithelium of the cornea of a 91-year-old woman transformed with the early region of the SV40 genome including genes encoding large T-antigen and small t-antigen; the subclone is described in the literature to represent differentiated cells of the so-called corneal endothelium |
Reference(s): | 15035, 14376, 18145 |
Biosafety level: | 1, GMO-S1 |
Permissions and restrictions: | A, D |
Depositor-specific restrictions: | Transfer to third parties needs to be negotiated with the depositor. |
| DSMZ Cell Culture Data: |
Morphology: | cuboid epitheloid cells growing strongly adherent in monolayers; image; image; image; image |
Medium: | Human-Endothelial-SFM + 10 ng/ml FGF-2 (observe the shelf-life of the medium). Do not apply FBS in order to keep differentiation status |
Subculture: | seed out at ca. 1 x 106 cells/80 cm2 flask; flasks must be coated with 10 µg/ml laminin and 10 mg/ml chondroitin sulfate; split confluent culture ca. 1:3 every 3-4 days using 2-3 ml trypsin/EDTA per 80 cm2 flask for 2-5 min at 37 °C (wash cells twice with PBS before trypsination, avoid trypsination longer than 5 min and check cell dissociation by microscopy); stop trypsination by using medium containing protease inhibitor cocktail (Sigma P1860) at 500-fold dilution; do not apply antibiotics, cells are highly sensitive to antibiotics |
Incubation: | at 37 °C with 5% CO2 |
Doubling time: | ca. 40-50 hours |
Harvest: | cell harvest of ca. 15 x 106 cells/175 cm2 |
Storage: | frozen with 90% medium (including FGF-2), 10% DMSO |
| DSMZ Scientific Data: |
Mycoplasma: | negative in microbiological culture, PCR assays |
Immunology: | To inquire about expression of EpCAM and intermediate filaments, contact ulfert.rand@dsmz.de. |
Authenticity: | STR analysis according to the global standard ANSI/ATCC ASN-0002.1-2021 (2021) resulted in an authentic STR profile of the reference STR database, matching the autologous cell line HCEC-12 |
Viruses: | PCR: EBV -, HBV -, HCV -, HIV-1 -, HIV-2 -, HTLV-1/2 -, MLV -, SMRV - |
Supplied as: | Delivery form | | Prices | | Frozen culture | | 450,- € | Growing culture (please inquire for exact delivery time) | | 900,- € | DNA isolated from cell line (25 µg) | | 560,- € | DNA isolated from cell line (5 µg) | | 140,- € | see price list |
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