Summary
The potential of cell and gene therapy has generated extensive interest over the past several years. More recently, identification of stem cells of various types, especially embryonic stem cells, reinforced this interest. Systematic studies are now being launched to define the biology of various stem cells, including after transplantation of cells in immunodeficient animals. This requires robust and unequivocal means to identify transplanted cells. Ideally, it should be possible to screen animal tissues for human cells with relatively simpler methods, followed by more precise localization of transplanted cells. We describe the application of conserved primate Charcot-Marie-Tooth disease type 1A repeat element for polymerase chain reaction-based screening of animal tissues for human cells. Similarly, direct polymerase chain reaction labeling of pancentromeric human alphoid sequences with digoxigenin-UTP generates in situ hybridization probes for identifying transplanted human cells. This pancentromeric probe identifies human cells irrespective of the original tissue source and can be combined with additional in situ methods to analyze cell differentiation. Incorporation of these strategies will facilitate translational studies aimed at understanding mechanisms concerning the trafficking, engraftment, proliferation, differentiation and function of human stem cells in animals.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Gupta S. and Chowdhury J. R. (2002) Therapeutic potential of hepatocyte transplantation. Semin. Cell Dev. Biol. 13, 439–446.
Malhi H., Irani A.N., Gagandeep S., and Gupta S. (2002) Isolation of human progenitor liver epithelial cells with extensive replication capacity and differentiation into mature hepatocytes. J. Cell Sci. 115, 2679–2688.
Brown J. J., Parashar B., Moshage H., et al. (2000) Long-term hepatitis B viremia model generated by transplanting nontumorigenic immortalized human hepatocytes in Rag-2-deficient mice. Hepatology 31, 173–181.
Dandri M., Burda M. R., Torok E., et al. (2001) Repopulation of mouse liver with human hepatocytes and in vivo infection with hepatitis B virus. Hepatology 33, 981–988.
Mercer D. F., Schiller D. E., Elliott J. F., et al. (2001) Hepatitis C virus replication in mice with chimeric human livers. Nat. Med. 7, 927–933.
Eaves C., Jiang X., Eisterer W., et al. (2003) New models to investigate mechanisms of disease genesis from primitive BCR-ABL(+) hematopoietic cells. Ann. N. Y. Acad. Sci. 996, 1–9.
Gupta S. and Rogler C. E. (1999) Lessons From Genetically Engineered Animal Models VI. Liver repopulation systems and study of pathophysiological mechanisms in animals. Am. J. Physiol. 277, G1097–G1102.
Follenzi A., and Gupta S. (2004) The promise of lentiviral gene therapy for liver cancer. J. Hepatol. 40, 337–340.
Cho J. J., Malhi H., Wang R., et al. (2002) Enzymatically labeled chromosomal probes for in situ identification of human cells in xenogeneic transplant models. Nat. Med. 8, 1033–1036.
Keller M. P., Seifried B. A., and Chance P. F. (1999) Molecular evolution of the CMT1A-REP region: a human-and chimpanzee-specific repeat. Mol. Biol. Evol. 16, 1019–1026.
Kiyosawa H. and Chance P. F. (1996) Primate origin of the CMT1A-REP repeat and analysis of a putative transposon-associated recombinational hotspot. Hum. Mol. Genet. 5, 745–753.
Baldini A., Ried T., Shridhar V., et al. (1993) An alphoid DNA sequence conserved in all human and great ape chromosomes: evidence for ancient centromeric sequences at human chromosomal regions 2q21 and 9q13. Hum. Genet. 90, 577–583.
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Benten, D., Cheng, K., Gupta, S. (2006). Identification of Transplanted Human Cells in Animal Tissues. In: Darby, I.A., Hewitson, T.D. (eds) In Situ Hybridization Protocols. Methods in Molecular Biology™, vol 326. Humana Press. https://doi.org/10.1385/1-59745-007-3:189
Download citation
DOI: https://doi.org/10.1385/1-59745-007-3:189
Publisher Name: Humana Press
Print ISBN: 978-1-58829-402-9
Online ISBN: 978-1-59745-007-2
eBook Packages: Springer Protocols