3D-Printed Collagen Scaffolds Promote Maintenance of Cryopreserved Patients-Derived Melanoma Explants

Abstract

The development of an in vitro three-dimensional (3D) culture system with cryopreserved biospecimens could accelerate experimental research screening anticancer drugs, potentially reducing costs and time bench-to-beside. However, minimal research has explored the application of 3D bioprinting-based in vitro cancer models to cryopreserved biospecimens derived from patients with advanced melanoma. We investigated whether 3D-printed collagen scaffolds enable the propagation and maintenance of patient-derived melanoma explants (PDMEs). 3D-printed collagen scaffolds were fabricated with a 3DX bioprinter. After thawing, fragments from cryopreserved PDMEs (approximately 1–2 mm) were seeded onto the 3D-printed collagen scaffolds, and incubated for 7 to 21 days. The survival rate was determined with MTT and live and dead assays. Western blot analysis and immunohistochemistry staining was used to express the function of cryopreserved PDMEs. The results show that 3D-printed collagen scaffolds could improve the maintenance and survival rate of cryopreserved PDME more than 2D culture. MITF, Mel A, and S100 are well-known melanoma biomarkers. In agreement with these observations, 3D-printed collagen scaffolds retained the expression of melanoma biomarkers in cryopreserved PDME for 21 days. Our findings provide insight into the application of 3D-printed collagen scaffolds for closely mimicking the 3D architecture of melanoma and its microenvironment using cryopreserved biospecimens.

Perspectives for 3D-Bioprinting in Modeling of Tumor Immune Evasion

“…(A) (B) Although 3D-bioprinting has been used in studies focusing on various forms of solid tumors, including neuroblastoma [66,67], melanoma [68,69], pancreatic cancer [55,70], non-small cell lung cancer [71], liver cancer [72], and osteosarcoma [73], the most prolific research has been conducted in breast cancer and glioma/glioblastoma models. The level of control over the spatial arrangement of cells, the possibility of multiple cell type use, and fine-tuning the biochemical and physical properties of biomaterials available through bioprinting allows generation of structures of a varied degree of complexity-from simple monocellular models to organoids and assembloids.…”

“…Although 3D-bioprinting has been used in studies focusing on various forms of solid tumors, including neuroblastoma [66,67], melanoma [68,69], pancreatic cancer [55,70], non-small cell lung cancer [71], liver cancer [72], and osteosarcoma [73], the most prolific research has been conducted in breast cancer and glioma/glioblastoma models.…”

3D Bioprinting: An Enabling Technology to Understand Melanoma

“…Such models are appealing and urgently needed as platforms for the screening and validation of drugs and cosmetics, as well as to perform fundamental studies to unravel mechanisms underlying skin diseases. Efforts have also been made in creating 3D bioprinted models of skin diseases, such as atopic dermatitis [ 88 ], psoriasis [ 156 ], and cutaneous melanoma [ 157 , 158 ]. However, the reported achievements in bioprinting diseased skin models are more modest so far.…”

“…In a subsequent study, a more detailed characterisation was performed to correlate the properties (e.g., mechanical properties) of hydrogels (alginate, alginate dialdehyde crosslinked with gelatin, and thiol-modified hyaluronan crosslinked with polyethylene glycol diacrylate) with the response of melanoma cells, but using a manual approach to create the hydrogels instead of bioprinting [ 213 ]. Using a different strategy, extrusion-based bioprinting was explored to create collagen scaffolds to support the maintenance and survival of cryopreserved patient-derived melanoma explants seeded onto the bioprinted scaffolds ( Figure 7 B) [ 158 ]. The results showed improved cell maintenance and survival compared to standard 2D culture and retention of key melanoma biomarker expression.…”

Instructive Hydrogels for Primary Tumor Cell Culture: Current Status and Outlook

“…[ 32 ] Surgically excised tissue is widely used to isolate primary tumor cells for the most of solid tumors. [ 26,33 ] To treat patients suffered from breast, [ 23 ] liver, [ 25 ] melanoma, [ 34 ] gastric, or colon cancer, [ 30 ] surgery is usually needed to remove malignant lesions from the body. These surgical resections provide a large number of primary tumor cells to ensure the successful establishment of PTOs in vitro.…”