Abstract
There is growing interest in developing drugs that specifically target glioblastoma tumor-initiating cells (TICs). Current cell culture methods, however, cannot cost-effectively produce the large numbers of glioblastoma TICs required for drug discovery and development. In this paper we report a new method that encapsulates patient-derived primary glioblastoma TICs and grows them in 3 dimension thermoreversible hydrogels. Our method allows long-term culture (~50 days, 10 passages tested, accumulative ~>1010-fold expansion) with both high growth rate (~20-fold expansion/7 days) and high volumetric yield (~2.0 × 107 cells/ml) without the loss of stemness. The scalable method can be used to produce sufficient, affordable glioblastoma TICs for drug discovery.
Highlights
Primary glioblastoma tumor-initiating cells (TICs) have been successfully isolated and cultured for long term, maintaining their capability for self-renewing[1,5,6,7,8,9,10]
We first confirmed the literature result that glioblastoma TICs could be cultured as 2 dimension (2D) adherent monolayer[1,5]
Cells could be propagated for multiple passages (10 passages tested in our laboratory) without significant differentiation as shown by the expression of glioblastoma TICs marker, Nestin, in the majority of cells (Fig. S1b,d)
Summary
Primary glioblastoma TICs have been successfully isolated and cultured for long term, maintaining their capability for self-renewing[1,5,6,7,8,9,10]. Glioblastoma TICs can form tumors with structures similar to the primary tumors These cultured glioblastoma TICs are invaluable for developing new drugs that can induce their death or differentiation, or sensitivity to current therapies. Glioblastoma TICs are either cultured as 2 dimension (2D) adherent monolayer or as 3 dimension (3D) neurospheres[1,5,6,7,8,9,10] While these methods can generate sufficient cells for basic science research, both are limited in their ability to produce large numbers of cells required for drug discovery and screening. A neurosphere culture method requires tens of liter volume to produce sufficient cells to screen million-compound library one time leading to the high cost for drug development. We systematically compared this new method with the 2D monolayer culture and the 3D neurosphere culture
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