Improving chimaerism quantification in bone marrow transplant recipients by image processing and analysis after restriction endonuclease in situ digestion (IPA-REISD).
Restriction endonuclease in situ digestion (REISD) with Sau3A of human metaphase chromosomes and interphase nuclei produces a conspicuous banding pattern involving pericentromeric regions of chromosomes 9 and 3. Constitutive heterochromatin of chromosome 9 is never digested by this enzyme while that of chromosome 3 is polymorphic, giving rise to three possible karyotypes: homozygous digested (3--), homozygous undigested (3++) or heterozygous individuals (3+-). Discrimination of this polymorphism between donor and recipient cells constitutes a rapid sex-independent method to monitor quantitatively the chimaerism achieved after bone marrow transplantation. An image processing and analysis (IPA)-assisted procedure which resolves residual fluorescent regions in metaphase chromosomes or interphase nuclei after REISD has been developed. IPA-REISD has interesting advantages over the basic REISD method by allowing a rapid, objective and precise discrimination of the polymorphism in large cell samples.
Buño
I
I
Unidad de Genetica, Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, Spain.
López-Fernández
C
C
Fernández
J L
JL
Llamas
P
P
Díez-Martin
J L
JL
Gosálvez
J
J
D001853:Bone Marrow / Q000473:pathology; D016026:Bone Marrow Transplantation; D002893:Chromosomes, Human, Pair 3; D002899:Chromosomes, Human, Pair 9; D004262:DNA Restriction Enzymes; D005260:Female; D006801:Humans; D007091:Image Processing, Computer-Assisted; D017403:In Situ Hybridization; D007399:Interphase; D007938:Leukemia / Q000235:genetics / Q000473:pathology / Q000628:therapy; D008297:Male; D008677:Metaphase; D008856:Microscopy, Fluorescence; D011110:Polymorphism, Genetic; D018183:Transplantation Chimera / Q000235:genetics*; D014184:Transplantation, Homologous
D004262:DNA Restriction Enzymes