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NKIM-6, a new immortalized human brain capillary endothelial cell line with conserved endothelial characteristics

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Abstract

Primary human brain capillary endothelial cells (hBCECs) are available only in small quantities and have a short life span in vitro; this restricts their use as in vitro model for the blood-brain barrier (BBB). To overcome these limitations, we have established an immortalized hBCEC line (NKIM-6) by transfection with pLXSN16E6E7, which encodes the human papillomavirus type 16 E6 and E7 genes. The cell line exhibits an extended life span in vitro and retains its characteristic endothelial morphology, endothelial markers, and physiology. Likewise, as demonstrated by immunohistochemistry and reverse transcription/polymerase chain reaction (RT-PCR), NKIM-6 cells express BBB markers, and the lack of glial, neuronal, and epithelial markers confirms their endothelial origin. Moreover, with quantitative RT-PCR, we have been able to demonstrate that several ATP-binding cassette-transporters are expressed in NKIM-6 cells with a conserved expression order compared with primary hBCECs. Our results suggest that this cell line might be suitable as in vitro model for several aspects of the BBB.

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Acknowledgements

The authors thank Corina Mueller, Stephanie Rosenzweig, Jutta Kocher, and Renate Steinle for excellent technical assistance, Dr. Heike Lindenmaier for an introduction into the use of the confocal laser-scanning microscope, Dr. Hans-Herbert Steiner (Department of Neurosurgery, University of Heidelberg, Germany) for providing tissue, and Dr. Horst-Peter Schmitt (Department of Neuropathology, University of Heidelberg, Germany) for histological evaluation of tissue. The cell lines MDCK I/MRP1, MDCK II/MRP2, and MDCK II/MRP3 were generously provided by Dr. Piet Borst (The Netherlands Cancer Institute, Amsterdam, The Netherlands). HUVECs were a kind gift from Dr. Thomas Dengler (Department of Internal Medicine III, Cardiology, University of Heidelberg, Germany).

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Author notes

  1. Sandra Caldeira

    Present address: European Molecular Biology Organization, Meyerhofstrasse 1, 69117, Heidelberg, Germany

  2. Massimo Tommasino

    Present address: Unit of Infection and Cancer, International Agency for Research on Cancer, World Health Organization, 69372, Lyon, France

Authors and Affiliations

  1. Department of Internal Medicine VI, Clinical Pharmacology and Pharmacoepidemiology, University of Heidelberg, Im Neuenheimer Feld 410, 69120, Heidelberg, Germany

    Nahal Ketabi-Kiyanvash, Walter E. Haefeli & Johanna Weiss

  2. Department of Neurosurgery, University of Heidelberg, Im Neuenheimer Feld 400, 69120, Heidelberg, Germany

    Christel Herold-Mende & Farzaneh Kashfi

  3. Tumor Virology, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120, Heidelberg, Germany

    Sandra Caldeira & Massimo Tommasino

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  1. Nahal Ketabi-Kiyanvash
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  2. Christel Herold-Mende
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  3. Farzaneh Kashfi
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Correspondence to Johanna Weiss.

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This work was supported by the Friedrich-Fischer Nachlass, University of Heidelberg, Germany.

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Ketabi-Kiyanvash, N., Herold-Mende, C., Kashfi, F. et al. NKIM-6, a new immortalized human brain capillary endothelial cell line with conserved endothelial characteristics. Cell Tissue Res 328, 19–29 (2007). https://doi.org/10.1007/s00441-006-0348-4

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