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Formation of functional tissue from transplanted adrenocortical cells expressing telomerase reverse transcriptase

Nature Biotechnology volume 18, pages 39–42 (2000)Cite this article

Abstract

We report the first use of human telomerase reverse transcriptase (hTERT) expression in experimental xenotransplantation. Previously, we showed that bovine adrenocortical cells can be transplanted into severe combined immunodeficient (SCID) mice, and that these cells form functional tissue that replaces the animals' own adrenal glands. We cotransfected primary bovine adrenocortical cells with plasmids encoding hTERT, SV40 T antigen, neo, and green fluorescent protein. These clones do not undergo loss of telomeric DNA and appear to be immortalized. Two clones were transplanted beneath the kidney capsule of SCID mice. Animals that received cell transplants survived indefinitely despite adrenalectomy. The mouse glucocorticoid, corticosterone, was replaced by the bovine glucocorticoid, cortisol, in the plasma of these animals. The tissue formed from the transplanted cells resembled that formed by transplantation of cells that were not genetically modified and was similar to normal bovine adrenal cortex. The proliferation rate in tissues formed from these clones was low and there were no indications of malignant transformation.

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Figure 1: Telomerase activity by TRAP assay in bovine adrenocortical cells expressing human telomerase reverse transcriptase (hTERT) and in control clones.
Figure 2: Telomere restriction fragment analysis in hTERT-modified bovine adrenocortical cells and in control cells.
Figure 3: Levels of plasma cortisol in mice with transplanted cells.
Figure 4: Histological appearance of hematoxylin-eosin stained tissues formed by transplantation of hTERT-modified bovine adrenocortical cell clones and normal adrenocortical tissue.
Figure 5: Proliferation in tissues formed by transplantation of hTERT-modified bovine adrenocortical cell clones assessed by expression of Ki-67 antigen.

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Acknowledgements

This work was supported by grants from the National Institute on Aging (AG 12287 and AG 13663), the Army Materiel and Medical Command, and the Texas Advanced Technology Program. We thank Edna Wright for performing radioimmunoassays.

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  1. Huffington Center on Aging and Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, 77030, TX

    Michael Thomas, Lianqing Yang & Peter J. Hornsby

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  1. Michael Thomas
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  2. Lianqing Yang
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  3. Peter J. Hornsby
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Correspondence to Peter J. Hornsby.

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Thomas, M., Yang, L. & Hornsby, P. Formation of functional tissue from transplanted adrenocortical cells expressing telomerase reverse transcriptase. Nat Biotechnol 18, 39–42 (2000). https://doi.org/10.1038/71894

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