Bioprocessing of human glioblastoma brain cancer tissue.
Solid cancer tumors are thought to arise from aberrant stem cell populations, called cancer stem cells (CSCs). Hence, the development of effective cancer therapies may rely on developing methods that specifically target these cells. However, the scarcity of CSCs in vivo represents a major impediment to such research, as there is an insufficient supply for basic biochemical and genetic analyses. It is therefore necessary to develop methods to expand reproducibly CSC tissue in vitro in a controlled environment. To date, we have developed bioreactor protocols for the suspension culture of an aggressive and deadly type of brain cancer called glioblastoma multiforme (GBM). Human GBM-derived cells achieved a maximum cell density of 2.4 x 10(6) cells/mL after 24 days under high shear conditions in batch culture conditions. In comparison, fed-batch cultures achieved 4.5 x 10(6) cells/mL after 32 days. Characterization of bioreactor-expanded cells using both flow cytometry and a differentiation assay indicated that bioreactor-generated human GBM-derived cells have similar characteristics to the initial cell population and achieve >90% CD133 expression. Additionally, genomic characterization indicated that a very small number of key genes were differentially expressed in the bioreactor-expanded GBM-derived cells, thereby conserving the basic nature of the brain cancer tissue in the cell expansion process.
Panchalingam
Krishna M
KM
Schulich School of Engineering, University of Calgary , Calgary, Canada .
Paramchuk
Wendy J
WJ
Chiang
Chun-Yi Katherine
CY
Shah
Nameeta
N
Madan
Anup
A
Hood
Leroy
L
Foltz
Greg
G
Behie
Leo A
LA
D000071916:AC133 Antigen; D015703:Antigens, CD / Q000378:metabolism; D019149:Bioreactors; D001932:Brain Neoplasms / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D002449:Cell Aggregation; D002452:Cell Count; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D003470:Culture Media; D015870:Gene Expression; D005909:Glioblastoma / Q000235:genetics / Q000276:immunology / Q000473:pathology*; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D014411:Neoplastic Stem Cells / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D020411:Oligonucleotide Array Sequence Analysis; D010455:Peptides / Q000378:metabolism; D046509:Tissue Culture Techniques / Q000379:methods; D023822:Tissue Engineering / Q000379:methods*
D000071916:AC133 Antigen; D015703:Antigens, CD; D003470:Culture Media; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides