High Quality Multicellular Tumor Spheroid Induction Platform Based on Anisotropic Magnetic Hydrogel

In recent years, multicellular spheroid (MCS) culture has been extensively studied both in fundamental research and application fields since it inherits much more characteristics from in vivo solid tumor than conventional two-dimensional (2D) cell culture. However, anticell adhesive MCS culture systems such as hanging drop allow certain cell lines only to form loose, irregular aggregates rather than MCS with physiological barriers and pathophysiological gradients, which failed to mimic in vivo solid tumor in these aspects. To address this issue, we improved our previously established anisotropic magnetic hydrogel platform, enabling it to generate multicellular spheroids with higher efficiency. The qualities of multicellular tumor spheroids (MCTSs) obtained on our platform and from classic 3D culture systems were compared in terms of morphology, biological molecule expression profiles, and drug resistance. In this novel platform, mature MCTSs with necrotic cores could be observed in 1 week. And results of molecular biological assays with real time-PCR and western-blot confirmed that MCTSs obtained from our platform performed higher cell pluripotency than those obtained from the hanging drop system. Moreover, a lower cell apoptosis ratio and better viability of cancer cells were observed on our platform both under culturing and drug treatment. In conclusion, higher quality of MCTSs obtained from this anisotropic magnetic hydrogel than classic hanging drop system validate its potential to be an in vitro platform of inducing tumor MCTS formation and drug efficacy evaluation.