[Molecular identification of human/goat xenogeneic model].
OBJECTIVE
To identify the human hematopoietic stem cells from the human/goat xenogeneic model with molecular techniques.


METHODS
DNA and total RNA were extracted from 11 transplanted goat peripheral blood cells. Human CD(34), GPA and SRY genes were amplified with PCR in these samples, and CD(34), GPA mRNA transcripts were detected using RT-PCR in 5 and 6 goat peripheral blood cells, respectively. Southern blot analysis was performed in 8 goat DNAs to detect the human specific alpha-satellite sequence. Meanwhile FISH was also performed to detect the human cells in goat blood with a probe of human Y chromosome.


RESULTS
Human CD(34) and GPA genes could be detected with PCR in all the 11 goats, and SRY gene did in 5 goats transplanted with hematopoietic stem cells derived from male human babies. Southern blot showed that human specific alpha-satellite sequence was present in 8 goats. By RT-PCR, human CD(34) mRNA was detected in 5 experimental goats, GPA mRNA was found in the other 6 experimental goats and FISH assay showed that some peripheral blood cells of the human/goat xenogeneic model were positive.


CONCLUSIONS
Existence of human cells in the recipient goats was identified by molecular detection, which was feasible for the examination of human/goat xenogeneic models.
Chen
Meijue
M
Shanghai Institute of Medical Genetics, Shanghai Children Hospital, Shanghai 200040, China.
Yan
Jingbin
J
Fang
Yudan
Y
Ren
Zhaorui
Z
Xiao
Yanping
Y
Huang
Shuzhen
S
D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics; D015139:Blotting, Southern; D005260:Female; D024282:Genes, sry / Q000235:genetics; D006021:Glycophorins / Q000235:genetics; D006041:Goats; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D016133:Polymerase Chain Reaction; D018183:Transplantation Chimera / Q000235:genetics*; D014183:Transplantation, Heterologous
D018952:Antigens, CD34; D006021:Glycophorins