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HK-2

CRL-2190

HK-2 cells are derived from a normal, human adult male kidney. The cell line has applications in toxicology research.
Product category
Human cells
Organism
Homo sapiens, human
Morphology
epithelial
Tissue
kidney; Cortex; Proximal tubule
Disease
Papilloma
Applications
3D cell culture
High-throughput screening
Toxicology
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

Biosafety Icon Biosafety Level 2

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

Cells contain Human papillomavirus type 16 (HPV-16) sequences

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Detailed product information

Characteristics

Growth properties
Adherent
Derivation
HK-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.

Age
adult
Gender
Male
Immortalization method
HPV-16 transfected
Genes expressed
alkaline phosphatase; gamma glutamyltranspeptidase; leucine aminopeptidase; acid phosphatase; cytokeratin; alpha 3, beta 1 integrin; fibronectin
Expression markers
Epidermal growth factor (EGF), expressed
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.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete 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.
  • Temperature
    37°C
    Atmosphere
    95% Air, 5% CO2
    Handling procedure

    To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

    1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).
    2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
    3. Transfer the vial contents to a centrifuge tube containing  9.0 mL complete culture medium. and spin at approximately 125 x g for 5 to 7 minutes.
    4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a 25 cm2 culture flask.  It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
    5. Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product.
    Subculturing procedure
    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: Cell growth is dependent on epidermal growth factor.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°C 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°C.
    Subcultivation Ratio: A subcultivation ratio of 1:4 is recommended
    Medium Renewal: Every 2 to 3 days
    Reagents for cryopreservation
    Complete growth medium supplemented with 7.5% (v/v) DMSO (ATCC 4-X)

    Quality control specifications

    Mycoplasma contamination
    Not detected
    STR profiling
    Amelogenin: X,Y
    CSF1PO: 13
    D13S317: 9
    D16S539: 11,12
    D5S818: 12
    D7S820: 10,11
    THO1: 9
    TPOX: 8,9
    vWA: 17,18

    History

    Deposited as
    Homo sapiens
    Depositors
    RA Zager
    Special collection
    NCRR Contract

    Legal disclaimers

    Intended use
    This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
    Warranty

    The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

    Disclaimers

    This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

    While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

    This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

    Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

    Permits & Restrictions

    Import Permit for the State of Hawaii

    If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

    MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

    References

    Curated Citations

    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

    Frequently Asked Questions

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