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.
Artigo
em Inglês
| 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:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Doadores de Tecidos
/
Veias
/
Medula Óssea
/
Reação em Cadeia da Polimerase
/
Transplante de Medula Óssea
/
Repetições de Microssatélites
/
Quimerismo
/
Citometria de Fluxo
Limite:
Animais
/
Humanos
Idioma:
Inglês
Revista:
Korean Journal of Hematology
Ano de publicação:
2007
Tipo de documento:
Artigo
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