Int. J. Dev. Biol. 52: 365 - 370 (2008)

https://doi.org/10.1387/ijdb.072362mm

Vol 52, Issue 4

Prolonged human/sheep cellular chimerism following transplantation of human hemopoietic stem cells into the ewe celomic cavity

Open Access | Short Communication | Published: 1 April 2008

Monica Michelini¹, Sandra Papini¹, Alfredo Rosellini¹, Giuseppe Noia², Maria S. Ligato², Salvatore Mancuso², Andrea Cavazzana³, Gloria Bertacca³, Claudio Di Cristofano³, Riccardo Saccardi⁴, Serena Urbani⁴ and Roberto P. Revoltella^1,*

¹Foundation O.N.L.U.S. "Stem Cells and Life", Pisa and Institute of Biomedical Technologies, C.N.R., Unit of Pisa, ²Department of Obstetric and Gynecology, Catholic University, Rome, ³Department of Oncology, Transplantation and Advanced Technology in Medicine, University of Pisa, Pisa and ⁴Bone Marrow Transplantation Unit, Careggi Hospital, Florence, Italy

Abstract

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-DQalpha1 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-DQalpha1 expression demonstrated multilineage engraftment. Both HLA-DQalpha1 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 HLADQalpha1 expression.

Keywords

cord blood, CD133+ cell, celomic cavity, transplantation, chimerism