Identification of transplanted human cells in animal tissues.
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 mmunodeficient 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.
Benten
Daniel
D
Department of Medicine, Albert Einstein College, Bronx, NY, USA.
Cheng
Kang
K
Gupta
Sanjeev
S
D000818:Animals; D015415:Biomarkers / Q000737:chemistry; D002503:Centromere / Q000737:chemistry; D004247:DNA / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D004076:Digoxigenin / Q000378:metabolism; D006801:Humans; D017403:In Situ Hybridization; D051379:Mice; D016133:Polymerase Chain Reaction; D033581:Stem Cell Transplantation; D020079:Terminal Repeat Sequences; D014183:Transplantation, Heterologous
D015415:Biomarkers; D015342:DNA Probes; D004247:DNA; D004076:Digoxigenin