Abstract

Background: Although a therapeutic effect of dendritic cell (DC)-based immunotherapy, a kind of regenerative medicine, has been recognized in various types of cancer including malignant glioma, it is still impractical because of several unsolved problems. This study is aimed to solve the problems in regenerative medicine through a clinical trial of immunotherapy using fusions of DCs and glioma cells (GCs) against malignant glioma, and to put it into practical use. Methods: Primary cultured GCs and glioma stem cells (GSCs) were generated from surgical specimens of patient. DCs were generated from PBMC of same patient, and were fused with GCs and GSCs. The entire process of cell production must be performed by pairs of two cell-culture operators in a dedicated cell processing facility. We developed a remote cell-observation system for reducing hands work of operators. As a project to establish a preservation method, cryopreservation of glioma tissues, GCs/GSCs, DCs and fusion cells followed by their viability examination. Results: The remote cell-observation system worked stable in morphological observation and cell-counting for adhesion cells. A growth curve was also automatically and accurately created. Although a morphological observation of floating cells such as GSCs and DCs was possible, there was some error in counting of those cells. A project to establish a preservation method is currently underway, including the development of storage containers and storage liquids. Conclusions: Although the remote cell-observation system required some modifications at the observation site, depth of focus, etc. for floating cells, there was no problem in accuracy for adhesion cells compared with operator’s observation. This system, which can be easily installed at low cost, seemed to be helpful for practical use of regenerative medical products including this therapy. We are working on a project to establish a stable transportation and preservation method for prevalence of this treatment.

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