Whole-organ tissue engineering: Decellularization and recellularization of three-dimensional matrix liver scaffolds

Abstract

To report the results of whole liver decellularization by two different methods. To present the results of grafting rat and sheep decellularized liver matrix (DLM) into the normal rat liver and compare natural cell seeding process in homo/xenograft of DLM. To compare the results of in vitro whole liver recellularization with rats' neonatal green fluorescent protein (GFP)-positive hepatic cells with outcomes of in vivo recellularization process. Whole liver of 8 rats and 4 sheep were resected and cannulated via the hepatic vein and perfused with sodium dodecyl sulfate (SDS) or Triton + SDS. Several examinations were performed to compare the efficacy of these two decellularization procedures. In vivo recellularization of sheep and rat DLMs was performed following transplantation of multiple pieces of both scaffolds in the subhepatic area of four rats. To compare the efficacy of different scaffolds in autologous cell seeding, biopsies of homograft and xenograft were assessed 8 weeks postoperatively. Whole DLMs of 4 rats were also recellularized in vitro by perfusion of rat's fetal GFP-positive hepatic cells with pulsatile bioreactor. Histological evaluation and enzymatic assay were performed for both in vivo and in vitro recellularized samples. The results of this study demonstrated that the triton method was a promising decellularization approach for preserving the three-dimensional structure of liver. In vitro recellularized DLMs were more similar to natural ones compared with in vivo recellularized livers. However, homografts showed better characteristics with more organized structure compared with xenografts. In vitro recellularization of liver scaffolds with autologous cells represents an attractive prospective for regeneration of liver as one of the most compound organs. In vivo cell seeding on the scaffold of the same species may have more satisfactory outcomes when compared with the results of xenotransplantation. This study theoretically may pave the road for in situ liver regeneration probably by implantation of homologous DLM or in vitro recellularized scaffolds into the diseased host liver.

Dual-Purpose Bioreactors to Monitor Noninvasive Physical and Biochemical Markers of Kidney and Liver Scaffold Recellularization

“…6B, day 7) in agreement with other reports using different sources of hepatocytes. 5,12 An increasing trend in overall HepG2 metabolic activity was observed, and resazurin reduction (and calculated cell number) was significantly higher after 5 ( p = 0.002) and 7 days ( p < 0.001) of perfusion culture compared to 3 days (Fig. 6C).…”

“…Decellularization of livers and kidneys through perfusion of detergent solutions through the vasculature has been well documented 3,4 -with technical feasibility demonstrated using both large animal and humanscale organs [5][6][7][8] as well as small animal organs for repopulation with stem or immature progenitor cells. 7,[9][10][11][12][13] The current challenges and limitations to organ and tissue engineering have shifted to the cellular repopulation phase of tissue development where donor cells are reintroduced into wholeorgan ECM scaffolds; however, very little detail is typically provided describing bioreactor construction despite the identification of specific design features that are critical to optimize scaffold recellularization and overall function.…”

Decellularized spleen matrix for reengineering functional hepatic-like tissue based on bone marrow mesenchymal stem cells

“…Studies have demonstrated that decellularized matrix based tissue regeneration is feasible for many hollow and parenchyma organs, such as bladder, 8 skin, 9 blood vessels, 10 heart, 11 lung, 12 kidney, 13 as well as liver. 14 Nonetheless, potential clinical application of DLM-based HLT may be plagued by the selection of scaffold source, considering that human derived DLM is also insufficient, and the application of xenogeneic DLM is hindered by potential risks of zoonosis and immunological rejection. Herein, in order to expand the scaffold donor pool, we have established the decellularized spleen matrix (DSM), which had homogeneous ECM with DLM, and presented comparable condition for hepatocyte cultivation.…”

Technology features of decellularization of human liver fragments as tissue-specific fine-grained matrix for cell-engineering liver construction

“…В литературе описаны успешные эксперимен-ты по децеллюляризации целой печени животных: крыс [4], кроликов [5], свиньи [6]. Также описан эксперимент по децеллюляризации левой латераль-ной доли, взятой у пациента с гемангиомой пече-ни [7].…”