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| 产品名称： | HCEC-B4G12 |
|---|---|
| 商品货号： | TS260032 |
| <strong>Cell line </strong>： | HCEC-B4G12 |
| DSMZ no. ： | TS260032 |
| 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 Caucasian 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 |
| <strong> </strong>： | DSMZ Scientific Data: |
| Morphology： | cuboid epitheloid cells growing strongly adherent in monolayers; image; image; image |
| Medium ： | Human-Endothelial-SFM + 10 ng/ml FGF-2 (observe the shelf-life of the medium; do not apply h.i. 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 |
| Mycoplasma ： | negative in microbiological culture, PCR assays |
| Immunology ： | To inquire about expression of EpCAM and intermediate filaments, contact hilmar.quentmeier@dsmz.de. |
| Fingerprint ： | fluorescent nonaplex PCR of short tandem repeat markers revealed a unique DNA profile (subclone of the cell line HCEC-12) |
| Viruses ： | PCR: EBV -, HBV -, HCV -, HIV-1 -, HIV-2 -, HTLV-I/II -, MLV -, SMRV - |