Simple immuno bead labeling of leukemic cell clusters cultured in methylcellulose.
In studies of in vitro leukemic clonogenic cells it is of importance to determine the cell lineage of individual clusters grown in culture. A method is described for the in-situ identification of leukemic cell clusters in methylcellulose cultures. Whole cultures are dried and incubated with various monoclonal antibodies, followed by incubation with beads coated with secondary antibody. Clusters containing antibody-positive cells are heavily labeled with beads which simplifies the recognition and scoring of clusters using normal light microscopy. This method has general applications and can also be used to identify normal myeloid and lymphoid clusters depending on the availability of lineage specific monoclonal antibodies.
Bol
S J
SJ
Department of Clinical Haematology and Oncology, Royal Children's Hospital, Melbourne, Australia.
Pick
M A
MA
Ekert
H
H
D001853:Bone Marrow / Q000473:pathology; D001854:Bone Marrow Cells; D002648:Child; D006801:Humans; D016130:Immunophenotyping; D008747:Methylcellulose; D014411:Neoplastic Stem Cells / Q000473:pathology / Q000502:physiology; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics / Q000473:pathology*; D014407:Tumor Cells, Cultured; D014410:Tumor Stem Cell Assay / Q000379:methods
D008747:Methylcellulose