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Cancer Stem Cells
Article first published online: 5 JAN 2009
Copyright © 2008 AlphaMed Press
Volume 27, Issue 1, pages 29–39, January 2009
How to Cite
Chong, Y.-K., Toh, T.-B., Zaiden, N., Poonepalli, A., Leong, S. H., Ong, C. E. L., Yu, Y., Tan, P. B., See, S.-J., Ng, W.-H., Ng, I., Hande, M. P., Kon, O. L., Ang, B.-T. and Tang, C. (2009), Cryopreservation of Neurospheres Derived from Human Glioblastoma Multiforme. STEM CELLS, 27: 29–39. doi: 10.1634/stemcells.2008-0009
Author contributions: Y.-K.C., T.-B.T., S.H.L., and O.L.K.: collection and assembly of data, data analysis and interpretation; N.Z. and A.P.: collection of data; C.E.L.O., Y.Y., P.B.T., and M.P.H.: collection and assembly of data; S.-J.S., W.-H.N., and I.N.: provision of study material and patients; B.-T.A.: conception and design, financial support, provision of study material and patients, collection and assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript; C.T.: conception and design, financial support, collection and assembly of data, data analysis and interpretation, manuscript writing, final approval of manuscript. Y.-K.C. and T.-B.T., B.T.A., and C.T. contributed equally to this work.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLSExpress October 9, 2008; available online without subscription through the open access option.
- Issue published online: 5 JAN 2009
- Article first published online: 5 JAN 2009
- Manuscript Accepted: 22 SEP 2008
- Manuscript Received: 6 JAN 2008
- Biomedical Research Council of Singapore
- Singapore Cancer Syndicate
- Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research. Grant Number: A∗STAR
- National Medical Research Council. Grant Numbers: NMRC/0910/2004, R-185-000-088-213
- Academic Research Fund
- Ministry of Education. Grant Number: R-185-000-153-112
- Cancer stem cell
Cancer stem cells have been shown to initiate and sustain tumor growth. In many instances, clinical material is limited, compounded by a lack of methods to preserve such cells at convenient time points. Although brain tumor-initiating cells grown in a spheroid manner have been shown to maintain their integrity through serial transplantation in immune-compromised animals, practically, it is not always possible to have access to animals of suitable ages to continuously maintain these cells. We therefore explored vitrification as a cryopreservation technique for brain tumor-initiating cells. Tumor neurospheres were derived from five patients with glioblastoma multiforme (GBM). Cryopreservation in 90% serum and 10% dimethyl sulfoxide yielded greatest viability and could be explored in future studies. Vitrification yielded cells that maintained self-renewal and multipotentiality properties. Karyotypic analyses confirmed the presence of GBM hallmarks. Upon implantation into NOD/SCID mice, our vitrified cells reformed glioma masses that could be serially transplanted. Transcriptome analysis showed that the vitrified and nonvitrified samples in either the stem-like or differentiated states clustered together, providing evidence that vitrification does not change the genotype of frozen cells. Upon induction of differentiation, the transcriptomes of vitrified cells associated with the original primary tumors, indicating that tumor stem-like cells are a genetically distinct population from the differentiated mass, underscoring the importance of working with the relevant tumor-initiating population. Our results demonstrate that vitrification of brain tumor-initiating cells preserves the biological phenotype and genetic profiles of the cells. This should facilitate the establishment of a repository of tumor-initiating cells for subsequent experimental designs. STEM CELLS2009;27:29–39