title: Quantification of human neural stem cell engraftments in rat brains using ERV-3 real-time PCR.
abstract: Few sensitive and reliable methods have been available for quantifying the number of transplanted human neural stem cells (hNSC) in the animal brain. To develop an accurate method for quantifying the number of hNSC incorporated in rat brain, we performed real-time PCR on hNSC-transplanted rat brains using a target sequence for ERV-3, which is an endogenous retrovirus present with a known copy number in all human cells, but not present in rodent cells. A standard curve was developed for known amount of different mixes of hNSC and rat fibroblasts, and test samples were prepared by manually incorporating variable, predefined numbers of hNSCs into rat brains. A cerebral rat hemisphere injected with 10(7) hNSC revealed 1.125% chimerism. Moreover, a linear correlation was found between hNSC numbers injected and their concentrations in the rat brain. In conclusion, the developed quantitative ERV-3 assay enables a simple, fast, and reproducible detection and quantitation of hNSC numbers in the rat brain.
authors.affiliation: Stroke and Neural Stem Cell Laboratory in Clinical Research Institute, Stem Cell Research Center, Department of Neurology, Seoul National University Hospital, Seoul, South Korea.
authors.forename: Soon-Tae
authors.initials: ST
authors.lastname: Lee
authors.forename: Kon
authors.initials: K
authors.lastname: Chu
authors.forename: Eun-Hee
authors.initials: EH
authors.lastname: Kim
authors.forename: Keun-Hwa
authors.initials: KH
authors.lastname: Jung
authors.forename: Kyung-Bok
authors.initials: KB
authors.lastname: Lee
authors.forename: Dong-In
authors.initials: DI
authors.lastname: Sinn
authors.forename: Seung U
authors.initials: SU
authors.lastname: Kim
authors.forename: Manho
authors.initials: M
authors.lastname: Kim
authors.forename: Jae-Kyu
authors.initials: JK
authors.lastname: Roh
mesh_terms: D000818:Animals; D015415:Biomarkers; D001921:Brain / Q000166:cytology* / Q000821:virology; D002452:Cell Count / Q000379:methods*; D020077:Endogenous Retroviruses / Q000235:genetics*; D006801:Humans; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000637:transplantation* / Q000821:virology; D051381:Rats; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D033581:Stem Cell Transplantation
chemical_list: D015415:Biomarkers