Open AccessCryopreservation of Neurospheres Derived from Human Glioblastoma Multiforme
Author(s) -
Chong YukKien,
Toh TanBoon,
Zaiden Norazean,
Poonepalli Anuradha,
Leong Siew Hong,
Ong Catherine Ee Ling,
Yu Yiting,
Tan Patrick B.,
See SiewJu,
Ng WaiHoe,
Ng Ivan,
Hande Manoor P.,
Kon Oi Lian,
Ang BengTi,
Tang Carol
Publication year - 2009
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2008-0009
Subject(s) - biology , neurosphere , cryopreservation , stem cell , population , glioma , olig2 , transplantation , brain tumor , vitrification , cancer research , cancer stem cell , transcriptome , regenerative medicine , immunology , andrology , microbiology and biotechnology , pathology , cellular differentiation , adult stem cell , oligodendrocyte , genetics , embryo , myelin , medicine , neuroscience , gene expression , gene , central nervous system , environmental health
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. S TEM C ELLS 2009;27:29–39