[Detection of human/goat xenogeneic models by interphase fluorescence in situ hybridization].
OBJECTIVE
To establish a high sensitive and specific method of interphase fluorescence in situ hybridization (IFISH) to detect the low-frequency human cells in human/goat xenogeneic models.


METHODS
Human-specific Y-chromosome satellite DNA CEPY and 17-chromosome satellite DNA p17H8 were used as probes for IFISH. The peripheral blood samples from 2 goats transplanted with human male hematopoietic stem cells (HSC), 1 normal negative goat and 1 normal man were analyzed. The actual FISH efficiency was confirmed by serial dilutions (1/100, 1/500 and 1/1000) of the cell mixture of normal man and normal negative goat. A set of signal scoring criteria was determined to guarantee the stability and reliability of the method.


RESULTS
Positive cell (human cell) frequencies were consistent with the established frequencies for the human/goat cell mixture. The average frequencies of positive cells were 98.60% (CEPY) and 100% (p17H8) for normal man, 0 for normal negative goat, 0.23% (CEPY) and 0.11% (p17H8) for human/goat xenogeneic models. The results demonstrated that low-frequency human cells (male cells confirmed by Y-chromosome probe) existed in human/goat xenogeneic models.


CONCLUSIONS
The IFISH developed in this study is of high sensitivity and specificity and can identify the actual frequency of human cells, which offers a direct, sensitive and specific approach to the detection of low-frequency human cells in human/goat xenogeneic models.
Xiao
Yanping
Y
Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai, 200040 P. R. China. sz.huang@verizon.net
Chen
Meiyue
M
Sheng
Min
M
Yang
Hua
H
Huang
Shuzhen
S
D000818:Animals; D002886:Chromosomes, Human, Pair 17 / Q000235:genetics; D041322:Chromosomes, Human, Y / Q000235:genetics; D015342:DNA Probes; D006041:Goats; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007399:Interphase / Q000235:genetics; D018895:Microsatellite Repeats / Q000235:genetics; D018183:Transplantation Chimera / Q000097:blood / Q000235:genetics*; D014183:Transplantation, Heterologous
D015342:DNA Probes