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UK Health Security Agency

HPCT-05-wt

HPCT-05-wt

Catalogue No.

12020703

Cell Line Name

HPCT-05-wt

Cell Line Description

HPCT-05-wt was developed from a primary cultures of proximal tubule epithelial cells from a 50-year old male which were infected with a replication-defective retroviral construct encoding wild-type simian virus 40 large T-antigen. Cells forming electrically resistive monolayers were selected and expanded in culture. HPCT-05-wt cells form polarized, resistive epithelial monolayers with apical microvilli, tight junctional complexes, numerous mitochondria, well-developed Golgi complexes, extensive endoplasmic reticulum, convolutions of the basolateral plasma membrane, and a primary cilium. The HPCT-05-wt cell line exhibits succinate, phosphate, and Na,K- adenosine triphosphatase (ATPase) transport activity, as well as acidic dipeptide- and adenosine triphosphate-regulated mechanisms of ion transport. Transcripts for Na(+)-bicarbonate cotransporter, Na(+)-H(+) exchanger isoform 3, Na,K-ATPase, parathyroid hormone receptor, epidermal growth factor receptor, and vasopressin V2 receptor were identified. Furthermore, immunoreactive sodium phosphate cotransporter type II, vasopressin receptor V1a, and CLIC-1 (NCC27) were also identified. The Y chromosome could not be detected in this cell line by short tandem repeat (STR)-PCR analysis when tested at ECACC. It is a known phenomenon that the Y chromosome can be rearranged or lost in cultured cells resulting in lack of detection. The cell line is identical to the source provided by the depositor based on the STR-PCR analysis. HPCT-05-wt will differentiate at confluence when cultured on a microporous support. The cells should be seeded at 105 cells/cm² on collagen-coated 30mm Millicell-CM culture plate inserts (0.4 µm, Millipore) at 37°C. EGF and serum should be omitted from the apical side of the chamber. On differentiation the cells form polarised resistive epithelial monolayers with apical microvilli, tight junctional complexes, numerous mitochondria, well-developed Golgi complexes, extensive endoplasmic reticulum, convolutions of the basolateral plasma membrane and primary cilium.

Characteristics

Tissue of Origin

Kidney

DNA profile (STR Profile)

Amelogenin: X
CSF1PO: 10
D5S818: 12,13
D7S820: 9,12
D13S317: 9
D16S539: 11,13
TH01: 6,9.3
TPOX: 8,10
vWA: 14,17

Disease

None Stated

Culture Conditions

Cell Type

Epithelial

Subculture Routine

Split subconfluent cultures (70-80%) 1:3 to 1:6 using 0.05% trypsin/EDTA; 5% CO₂; 37°C. Suggested seeding density 2-4x10,000 cells/cm². Saturation density is approximately 12 x 10,000 cells/cm². Flasks must be pre-coated with Collagen I (Sigma C3867) diluted 1:100 with medium without serum. Coating is applied overnight at 37°C then the flask rinsed with PBS prior to use.

Culture Medium

Stemline Keratinocyte Medium II (Sigma S0196) + Stemline Keratinocyte Growth Supplement (Sigma S9945) + 2mM Glutamine + 5 ng/ml human recombinant epidermal growth factor + 5% Foetal Bovine Serum (FBS). An alternative medium formulation can be found in Orosz et al., 2004 PMID: 14748622.

Growth Mode

Adherent

Additional Info

Depositor

Professor Ulrich Hopfer, Department of Physiology and Biiophysics, School of Medicine, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, Ohio 44106-4970

Country of Origin

United States

GMO Status

Genetically Modified Organism Class 1 (GMO1)

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

References

Orosz DE, Woost PG, Kolb RJ, Finesilver MB, Jin W, Frisa PS, Choo CK, Yau CF, Chan KW, Resnick MI, Douglas JG, Edwards JC, Jacobberger JW, Hopfer U. 2004 Growth, immortalization, and differentiation potential of normal adult human proximal tubule cells. In Vitro Cell Dev Biol Anim. 40(1-2):22-34. PMID: 14748622.

Bibliography

Jat PS, Cepko CL, Mulligan RC, Sharp PA. 1986 Recombinant retroviruses encoding simian virus 40 large T antigen and polyomavirus large and middle T antigens. Mol Cell Biol. 6(4):1204-17. PMID: 3023876. Woost PG, Kolb RJ, Finesilver M, Mackraj I, Imboden H, Coffman TM, Hopfer U. 2006 Strategy for the development of a matched set of transport-competent, angiotensin receptor-deficient proximal tubule cell lines. In Vitro Cell Dev Biol Anim. 42(7):189-200. PMID: 16948500.

Available Formats

  • Frozen
  • DNA-5µg (100ng/µl)

If use of this culture results in a scientific publication, it should be cited in the publication as: HPCT-05-wt (ECACC 12020703).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.