Prolonged human/sheep cellular chimerism following transplantation of human hemopoietic stem cells into the ewe celomic cavity.
We evaluated the possibility of prolonged chimerism formation in fetus and lamb, following human cord blood-selected CD133+ hemopoietic stem cell (HSC) transplantation into the celomic cavity of ewes at a pre-immune fetal age (44-45 days of pregnancy). Nineteen ewes were injected with HSC and 5 controls with a saline solution. By PCR, HLA-DQ alpha 1 and 6 human microsatellites (CODIS) were used for HSC traceability. FISH analysis was performed with 8 human DNA probes from different chromosomes, to confirm chromosomal integrity, nuclear DNA localization and donor DNA identification. Immunological staining for revealing HLA-DQ alpha 1 expression demonstrated multilineage engraftment. Both HLA-DQ alpha 1 and microsatellites were detected in different tissues of 3 available aborted fetuses, to a lesser extent in 11 lambs tested at 2-months, but not 12-months after birth. Although only 1 fetus of siblings of each sheep was injected, all siblings revealed positive engraftments. Microsatellite analysis showed evidence of human allele segregation in different tissues of individual fetuses and lambs. FISH analysis confirmed chimerism and the presence of human chromosomes. Non-detection of some human gene sequences in different chromosomes and random finding of allele segregation for some human heterozygous microsatellites were found in different tissues of individual animals. Controls born from un-transplanted ewes never revealed any human DNA sequences nor HLADQ alpha 1 expression.
Michelini
Monica
M
Foundation ONLUS Stem Cells and Life, Pisa and Institute of Biomedical Technologies, CNR, Unit of Pisa, Pisa, Italy.
Papini
Sandra
S
Rosellini
Alfredo
A
Noia
Giuseppe
G
Ligato
Maria S
MS
Mancuso
Salvatore
S
Cavazzana
Andrea
A
Bertacca
Gloria
G
Di Cristofano
Claudio
C
Saccardi
Riccardo
R
Urbani
Serena
S
Revoltella
Roberto P
RP
D000818:Animals; D001483:Base Sequence; D002877:Chromosomes, Human / Q000235:genetics; D036101:Cord Blood Stem Cell Transplantation / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005865:Gestational Age; D006085:Graft Survival / Q000235:genetics / Q000276:immunology; D006683:HLA-DQ Antigens / Q000378:metabolism; D059848:HLA-DQ alpha-Chains; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D018895:Microsatellite Repeats; D010529:Peritoneal Cavity; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D018183:Transplantation Chimera / Q000235:genetics / Q000276:immunology
D017931:DNA Primers; D006683:HLA-DQ Antigens; D059848:HLA-DQ alpha-Chains; C069050:HLA-DQA1 antigen