In utero transplantation of human hematopoetic stem cells into fetal goats under B-type ultrasonographic scan: an experimental model for the study of potential prenatal therapy.
OBJECTIVE
Using fetal goats as animal models, to establish the methodology of in utero transplantation of human hematopoeitic stem cell (HSC) under B-scan ultrasonographic guidance for prenatal therapy.


METHODS
Human HSC were directly injected into the peritoneal cavities of the recipient fetal goats at 45-55 days of gestation (term: 145 days) under the guidance of B-type ultrasound scan. After birth, the peripheral blood was collected for fluorescence assisted cell sorting (FACS), quantitative real-time PCR and fluorescence in situ hybridization (FISH) to detect and analyze the presence of human cells in the recipients.


RESULTS
The 32 recipients were born alive except one miscarriage. To test for the presence of human-goat chimeras, cells from 13 randomly selected transplanted goats were collected. FACS analyses showed the presence of human cells in all the transplanted goats tested. The average proportion of CD34+ cells and GPA+(glycophorin A) cells in the peripheral blood were 1.34 +/- 1.10% and 2.80 +/- 2.10%, respectively. No CD34+ or GPA+ cells were found in the non-transplanted goats tested. The results of the quantitative real-time PCR in three engraftment goats were 1.2 x 10(4), 2.9 x 10(4), and 3.2 x 10(4) copies of human GPA DNA per mug of genomic DNA. FISH experiments showed that cells containing human specific alpha-satellite DNA sequence were present in the peripheral blood of the transplanted goats.


CONCLUSIONS
The method described herein is safe and reliable, with low miscarriage risk and high chimerism rate. This approach may provide a promising animal model for potential prenatal treatment.
Zeng
Fanyi
F
Institute of Medical Genetics, Shanghai Children's Hospital, Shanghai Jiao Tong University, 24/1400 West Beijing Road, Shanghai 200040, PR China.
Chen
Mei-Jue
MJ
Huang
Wen-Ying
WY
Yan
Jing-Bin
JB
Xiao
Yan-Ping
YP
Gong
Zhi-Juan
ZJ
Ren
Zhao-Rui
ZR
Huang
Shu-Zhen
SZ
D000818:Animals; D002886:Chromosomes, Human, Pair 17 / Q000235:genetics; D004247:DNA / Q000097:blood; D004276:DNA, Satellite / Q000097:blood; D005260:Female; D005434:Flow Cytometry; D005865:Gestational Age; D006021:Glycophorins / Q000235:genetics; D006041:Goats / Q000097:blood / Q000196:embryology*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D023421:Models, Animal; D010529:Peritoneal Cavity / Q000196:embryology; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D033581:Stem Cell Transplantation / Q000379:methods*; D018183:Transplantation Chimera / Q000235:genetics; D014183:Transplantation, Heterologous; D014463:Ultrasonography / Q000379:methods*
D004276:DNA, Satellite; D006021:Glycophorins; D004247:DNA