Fast Automated Approach for the Derivation of Acellular Extracellular Matrix Scaffolds from Porcine Soft Tissues

Abstract

Decellularized extracellular matrix (ECM) scaffolds derived from tissues and organs are complex biomaterials used in clinical and research applications. A number of decellularization protocols have been described for ECM biomaterials derivation, each adapted to a particular tissue and use, restricting comparisons among materials. One of the major sources of variability in ECM products comes from the tissue source and animal age. Although this variability could be minimized using established tissue sources, other sources arise from the decellularization process itself. Overall, current protocols require manual work and are poorly standardized with regard to the choice of reagents, the order by which they are added, and exposure times. The combination of these factors adds variability affecting the uniformity of the final product between batches. Furthermore, each protocol needs to be optimized for each tissue and tissue source making tissue-to-tissue comparisons difficult. Automation and standardization of ECM scaffold development constitute a significant improvement to current biomanufacturing techniques but remains poorly explored. This study aimed to develop a biofabrication method for fast and automated derivation of raw material for ECM hydrogel production while preserving ECM composition and controlling lot-to-lot variability. The main result was a closed semibatch bioreactor system with automated dosing of decellularization reagents capable of deriving ECM material from pretreated soft tissues. The ECM was further processed into hydrogels to demonstrate gelation and cytocompatibility. This work presents a versatile, scalable, and automated platform for the rapid production of ECM scaffolds.

Fabrication of Size-Controllable and Arrangement-Orderly HepG2 Spheroids for Drug Screening via Decellularized Liver Matrix-Derived Micropattern Array Chips

“…Recent advances in whole organ and tissue decellularization have made it possible to obtain organ-specific ECM with unique architecture, composition, and biological and physical properties. − Compared with a single ECM protein component, such as collagen I which was widely used in the liver tissue engineering, liver-specific ECM containing various biomacromolecules directs signal molecules or interacts with tumor cells through integrins, discoidin domain receptors (DDRs), and transmembrane protein proteoglycans to regulate the signaling pathways related to cell proliferation, migration, and differentiation, thereby influencing the cells’ biological behavior. , …”

Ordered micropattern arrays fabricated by lung-derived dECM hydrogels for chemotherapeutic drug screening

“…Standardization is necessary before immortalized cancer cell lines can be applied in routine pharmacotoxicology. From organ sources and animal age, decellularized reagents and exposure times, to the shape, size and arrangement of cancer spheroids could strongly affect drug efficacy and toxicity [ 41 , 42 ]. The emergence of micropattern arrays will enhance the efficiency and affordability of the novel drug development process.…”

Towards a standardized multi-tissue decellularization protocol for the derivation of extracellular matrix materials

“…Standardized Tissue Decellularization All decellularizations were performed using an automated decellularization system previously described by our group. 24,25 Porcine VFLP, SG, heart, and lung tissues were procured from market weight pigs from Nahunta Pork Outlet (Raleigh, NC). Porcine skin, muscle, and tendon tissues were obtained from 3-6 months old pigs from the College of Veterinary Medicine (CVM) at North Carolina State University.…”

“…Previously, the combination of sodium deoxycholate, DNAse, peracetic acid, and DI water/DPBS washes was successfully applied to VFLP and SG. 24,25 Due to its success and significant prior data, this protocol represented the foundation for the six additional tissues decellularized in this study. Table 3 shows a summary explanation for why these Paper Biomaterials Science reagents were selected and other reagents that will be included in future studies.…”