A technique of bone marrow collection from vertebral bodies of cynomolgus macaques for transplant studies.
J Surg Res
; 124(2): 280-8, 2005 Apr.
Article
em En
| MEDLINE
| ID: mdl-15820259
ABSTRACT
BACKGROUND:
Strategies to induce donor-specific allograft tolerance are best tested in preclinical models developed in nonhuman primates (NHPs). Most protocols prepare the recipient by infusing hematopoietic cells from the donor. We report here a procedure to isolate and characterize large numbers of bone marrow cells (BMCs) from cynomolgus monkeys (cynos) that can then successfully be transplanted into conditioned recipients. MATERIALS ANDMETHODS:
Vertebral columns of five cynos were excised en bloc and separated into individual vertebrae. The cancelous bone was extracted with a core puncher, fractionated, filtered, centrifuged, and resuspended in transplantation media before being analyzed by flow cytometry. In two instances, the collected BMCs were reinfused into allogeneic recipients preconditioned with a nonmyeloablative regimen. Chimerism was monitored using short-tandem repeat analysis.RESULTS:
The mean total BMCs yield was 25.5 x 10(9) (range of 4.00 x 10(9) to 59 x 10(9)) with mean cell viability of 93.4% (range 90-96%). CD34+ cells and CD3+ cells averaged 0.34 and 3.91% of total BMCs, respectively. This resulted in absolute cell number yields of 1.02 x 10(8) and 1.15 x 10(9) for CD34+ and CD3+ cells, respectively. Graft-versus-host disease was absent in both bone marrow infused animals, and a maximum level of chimerism of 18% was detected at 3 weeks after BMCs infusion.CONCLUSION:
We present here the first detailed report of a procedure to retrieve and characterize large numbers of BMCs from vertebral bodies of cynos and demonstrate that cells collected with this technique have the capability of engrafting in allogenic recipients.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Coluna Vertebral
/
Células da Medula Óssea
/
Células-Tronco Hematopoéticas
/
Separação Celular
/
Transplante de Células-Tronco Hematopoéticas
Limite:
Animals
Idioma:
En
Ano de publicação:
2005
Tipo de documento:
Article