Practical Evaluation of Engraftment and Mixed Chimerism Using PCR Amplification of a Microsatellite in the Class II Eb Gene in Murine MHC-mismatched, Nonmyeloablative Bone Marrow Transplantation
Korean Journal of Hematology
;
: 91-97, 2007.
Artículo
en Inglés
| WPRIM
| ID: wpr-720132
ABSTRACT
BACKGROUND:
Although engraftment following murine allogeneic bone marrow transplantation (BMT) is most commonly confirmed by H2 typing using flow cytometry, recipient mice can be seriously injured during peripheral blood (PB) sampling. Therefore, we developed an alternative DNA-based assay that does not require the large volume of PB necessary for flow cytometry.METHODS:
A minute volume of PB from the tail vein was used to evaluate the engraftment by PCR amplification of a microsatellite in the class II Eb gene. Dilution experiments were performed to evaluate the sensitivity of this assay for detecting donor cells in mixed cell populations compared with flow cytometry analysis.RESULTS:
Early engraftment and mixed chimerism were confirmed, based on the length variation of the microsatellite in the class II Eb gene. The degree of donor chimerism in the donor-recipient cell mixture could be estimated semiquantitatively in a dilution experiment. The sensitivity of this assay by the naked eye approached 10% of the degree of donor chimerism.CONCLUSION:
PCR amplification of a microsatellite in the class II Eb gene can be a useful alternative to flow cytometry for evaluating early engraftment and mixed chimerism following murine nonmyeloablative BMT.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Donantes de Tejidos
/
Venas
/
Médula Ósea
/
Reacción en Cadena de la Polimerasa
/
Trasplante de Médula Ósea
/
Repeticiones de Microsatélite
/
Quimerismo
/
Citometría de Flujo
Límite:
Animales
/
Humanos
Idioma:
Inglés
Revista:
Korean Journal of Hematology
Año:
2007
Tipo del documento:
Artículo
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