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HK-2 拉丁名

(ATCC® CRL-2190™) 編號

Organism: Homo sapiens, human /

Cell Type: human papillomavirus 16 (HPV-16) transformed /

Tissue: kidney, cortex/proximal tubule /

Disease: Papilloma

Organism Homo sapiens, human

Tissue kidney, cortex/proximal tubule

Cell Type human papillomavirus 16 (HPV-16) transformed

Product Format 提供形式 frozen

Morphology epithelial Culture Properties adherent

Biosafety Level 生物安全等級 2 [Cells contain Papilloma viral DNA sequences]

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease Papilloma Age adult Gender male

Storage Conditions 保藏方法 liquid nitrogen vapor phase DerivationHK-2 (human kidney 2) is a proximal tubular cell (PTC) line derived from normal kidney.

The cells were immortalized by transduction with human papilloma virus 16 (HPV-16) E6/E7 genes.

The cell line appears to be derived from a single cell based on Southern and FISH analysis.

Clinical Data adult male Receptor Expressio epidermal growth factor (EGF), expressed Genes Expressed alkaline phosphatase;

gamma glutamyltranspeptidase; leucine aminopeptidase; acid phosphatase; cytokeratin; alpha 3, beta 1 integrin;

fibronectin Cellular Products alkaline phosphatase; gamma glutamyltranspeptidase; leucine aminopeptidase;

acid phosphatase; cytokeratin; alpha 3, beta 1 integrin; fibronectin

Comments 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 (see ATCC CRL-9078).

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.

Complete Growth Medium The base medium for this cell line is provided by Invitrogen (GIBCO) as part of a kit: Keratinocyte Serum Free Medium (K-SFM), Kit Catalog Number 17005-042.

This kit is supplied with each of the two additives required to grow this cell line (bovine pituitary extract (BPE) and human recombinant epidermal growth factor (EGF).

To make the complete growth medium, you will need to add the following components to the base medium: • 0.05 mg/ml BPE - provided with the K-SFM kit • 5 ng/ml EGF - provided with the K-SFM kit. NOTE: Do not filter complete medium.

Subculturing Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.

Note: The cells should not be allowed to become confluent, subculture at 80% of confluence.

1. Remove and discard culture medium.

2.Briefly rinse the cell layer with 0.05% (w/v) Trypsin- 0.53 mM EDTA solution.

3.Add 2.0 to 3.0 mL of 0.05% Trypsin-0.53 mM EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).

Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.

Cells that are difficult to detach may be placed at 37℃ to facilitate dispersal.

4.Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.

5.To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.

Discard supernatant and resuspend cells in fresh growth medium.

Add appropriate aliquots of cell suspension to new culture vessels.

6. Incubate cultures at 37℃.

Subcultivation Ratio: A subcultivation ratio of 1:4 is recommended Medium Renewal: Every 2 to 3 days Cryopreservation Freeze medium: Complete growth medium supplemented with 7.5% (v/v) DMSO

Storage temperature 保藏溫度 : liquid nitrogen vapor phase

Culture Conditions 培養(yǎng)條件 

Atmosphere 需氧情況 : air, 95%; carbon dioxide (CO2), 5%

Temperature 培養(yǎng)溫度 : 37℃

Growth Conditions 生長條件 : Cell growth is dependent on epidermal growth factor.

The cells should not be allowed to become confluent.

Subculture at 80% of confluence.

STR Profile Amelogenin: X,Y CSF1PO: 13 D13S317: 9 D16S539: 11,12 D5S818: 12 D7S820: 10,11 THO1: 9 TPOX: 8,9 vWA: 17,18

Name of Depositor RA Zager

References 參考文獻(xiàn) Ryan MJ, et al. HK-2: an immortalized proximal tubule epithelial cell line from normal adult human kidney. Kidney Int. 45: 48-57, 1994. PubMed: 8127021