Culture of Bladder Cancer Organoids as Precision Medicine Tools

GelMA, Click-Chemistry Gelatin and Bioprinted Polyethylene Glycol-Based Hydrogels as 3D Ex Vivo Drug Testing Platforms for Patient-Derived Breast Cancer Organoids

“…The tumor pieces were successfully reduced to individual cells, according to established protocols [ 40 ]. Relying on tissue mincing, enzymatic digestion and mechanical agitation, cells were then cultured and expanded in 2D ( Figure 1 A) using a suitable media validated for human BCa-derived organoids [ 41 ]. While it is acknowledged that this may lead to specific cell populations being selected, this strategy is needed to obtain sufficient cell numbers prior to 3D printing, as the surplus to pathology needs received for research purposes is too small.…”

“…While it is acknowledged that this may lead to specific cell populations being selected, this strategy is needed to obtain sufficient cell numbers prior to 3D printing, as the surplus to pathology needs received for research purposes is too small. The specific organoid media we used was mostly similar to the medium by Guillen et al and is recommended for BCa organoids [ 41 ]. It contains serum and specific growth factors, such as epidermal growth factor (EGF), that enhance epithelial cell proliferation, as well as the Rho kinase inhibitor (Y-27632), a critical supplement to support BCa ex vivo cultures past 15 days [ 41 ].…”

“…The specific organoid media we used was mostly similar to the medium by Guillen et al and is recommended for BCa organoids [ 41 ]. It contains serum and specific growth factors, such as epidermal growth factor (EGF), that enhance epithelial cell proliferation, as well as the Rho kinase inhibitor (Y-27632), a critical supplement to support BCa ex vivo cultures past 15 days [ 41 ].…”

“…This is in line with previous literature where peptide combination was more effective for organoid growth than their individual counterparts, due to higher number of adhesions sites [ 49 ]. For instance, incorporation of YGSIR in PEG hydrogels limited adhesion and no spreading of human microvascular endothelial cells and vascular smooth muscle cells, which was strongly enhanced when YGSIR was combined with RGD [ 41 ]. This suggests that desired response of specific cell types to PEG hydrogels could be optimized through the combinatory use of biomimetic peptides.…”

“…Surgically-resected tumor specimens were placed in a sterile Petri dish, minced to 0.5 × 1 mm 3 fragments using a sterile surgical scalpel and transferred to a sterile 50 mL centrifuge tube. A volume of 1 mL of 10× hyaluronase/collagenase (STEMCELL technologies, Tullamarine, VIC, Australia) and 4 mL of organoid media (ORM) [ 41 ] was added. ORM consisted of the following components supplied from Thermofisher, Brisbane, QLD, Australia; advanced DMEM/F12 (Gibco™, Waltham, MA, USA), supplemented with 10% ( v / v ) fetal bovine serum (FBS), 1% ( v / v ) penicillin streptomycin (P/S), HEPES 1M, GlutaMax 1×, Epidermal Growth Factor (EGF, 10 ng/mL).…”