Description
HK-2 Cell Line | CL-0109 | Gentaur US, UK & Europe Disrtribition
Media: CM-0109
Organism: Homo sapiens, Human
Application: N/A
Background: The recombinant retrovirus vector pLXSN 16 E6/E7 containing the HPV-16 E6/E7 genes was used to transfect the ectotropic packaging cell line Psi-2. Virus produced by the Psi-2 cells was used to infect the amphotropic packaging cell line PA317. Virus produced by the PA317 cells was used to transduce primary PTCs. Although pLXSN 16 E6/E7 also confers resistance to neomycin, selection in G418 was not used to isolate transduced clones. The cell line appears to be derived from a single cell based on Southern and FISH analysis. The E6/E7 genes are present in the HK-2 genome as determined by PCR. The cells retain a phenotype indicative of well differentiated PTCs. They are positive for alkaline phosphatase, gamma glutamyltranspeptidase, leucine aminopeptidase, acid phosphatase, cytokeratin, alpha 3,beta 1 integrin, and fibronectin. The cells are negative for factor Ⅷ related antigen, 6. 19 antigen and CALLA endopeptidase. HK-2 cells retain functional characteristics of proximal tubular epithelium such as Na＋dependent/phlorizin sensitive sugar transport and adenylate cyclase responsiveness to parathyroid, but not to antidiuretic hormone. The cells are capable of gluconeogenesis as evidenced by their ability to make and store glycogen. HK-2 cells are anchorage dependent. The cells will not grow in methylcellulose, soft agar or suspension. HK-2 cells can reproduce experimental results obtained with freshly isolated PTCs.
Age: Adult
Tissue: Kidney, cortex/proximal tubule
Morphology: Epithelial
Growth Properties: Adherent
Doubling Time: N/A
Biosafty: 2
Medium: MEM(PM150410)+10% FBS+1% P/S(PB180120)
Subculturing: Remove and discard culture medium. Briefly rinse the cell layer with DPBS solution to remove all traces of serum that contains trypsin inhibitor. Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 2 to 3 minutes). Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 4.0 to 6.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels.
Ratio: 1:2 to 1:4
Renewal: Every 2 to 3 days
Cryopreservation: Freeze medium: 60% Basal medium+30% FBS+10% DMSO Storage temperature: Liquid nitrogen vapor phase
Culture Conditions: Atmosphere: Air, 95%; CO2, 5% Temperature: 37℃
Effects: N/A
Duration: N/A
Lead Time: 3-5 days
