Laminin-adherent versus suspension-non-adherent cell culture conditions for the isolation of cancer stem cells in the DAOY medulloblastoma cell line.
Medulloblastoma (MB) is a highly malignant tumor of childhood. MB seems to be initiated and maintained by a small group of cells, known as cancer stem cells (CSCs). The CSC hypothesis suggests that a subset of tumor cells is able to proliferate, sustain the tumor, and develop chemoresistance, all of which make of CSC an interesting target for new anticancer therapies. The MB cell line DAOY was cultured in suspension by a medullosphere traditional culturing method and in adherent conditions by laminin-pre-coated flasks and serum-free medium enriched with specific growth factors. An increase in the stem features was shown when cells were successively cultured in hypoxia conditions. By contrast, a reduction in these properties was appreciated when cells were exposed to differentiation conditions. In addition, the CD133+ and CD133- subpopulations were isolated from cells grown in laminin-pre-coated flasks, and in vitro experiments showed that the CD133+ fraction represented the stem population and it could have CSC with a higher probability than the CD133- fraction. We can conclude that the laminin culture method in adherent conditions and the medullosphere traditional culturing method in suspension are similarly good for obtaining stem-like cells in the DAOY cell line.
Department of Biochemistry and Genetics, University of Navarra School of Sciences, Irunlarrea 1, Pamplona, 31008, Spain.
Javier
J
de la Rosa
Department of Biochemistry and Genetics, University of Navarra School of Sciences, Irunlarrea 1, Pamplona, 31008, Spain.
Ander
A
Sáenz Antoñanzas
Interdisciplinary Brain Research Centre, Faculty of Medicine, Aligarh Muslim University, Aligarh, India.
Mehdi H
MH
Shahi
Molecular Pathology Research Unit, Virgen de la Salud Hospital, Toledo, Spain.
Bárbara
B
Meléndez
IdiPaz Research Unit, La Paz University Hospital, Madrid, Spain.
Juan A
JA
Rey
Department of Biochemistry and Genetics, University of Navarra School of Sciences, Irunlarrea 1, Pamplona, 31008, Spain. jscastresana@unav.es.
Javier S
JS
Castresana
D000071916:AC133 Antigen / Q000235:genetics / Q000378:metabolism; D002448:Cell Adhesion / Q000235:genetics; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D002469:Cell Separation / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007797:Laminin / Q000378:metabolism*; D008527:Medulloblastoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018874:Spheroids, Cellular / Q000378:metabolism*; D014410:Tumor Stem Cell Assay
D000071916:AC133 Antigen; D007797:Laminin
CD133; Cancer stem cells; Hypoxia; Laminin; Medulloblastoma; Medullospheres; Neurospheres