HCEC-B4G12 cell line人角膜内皮细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

HCEC-B4G12 cell line人角膜内皮细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

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 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):
Valtink, M., Gruschwitz, R., Funk, R. H., Engelmann, K. (2008). Two clonal cell lines of immortalized human corneal endothelial cells show either differentiated or precursor cell characteristics. Cells Tissues Organs 187 (4): 286-294 . http://www.ncbi.nlm.nih.gov/pubmed/18196893
Biosafety level:
1, GMO-S1
Permissions and restrictions:
A, D

DSMZ Cell Culture Data:
Morphology:
cuboid epitheloid cells growing strongly adherent in monolayers;
Images:



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

DSMZ Scientific Data:
Mycoplasma:
negative in microbiological culture, PCR assays
Immunology:
To inquire about expression of EpCAM and intermediate filaments, contact BioVector NTCC Inc.
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 -

Supplier来源:BioVector NTCC Inc.
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