Decellularized Lymph Nodes as Scaffolds for Tissue Engineered Lymph Nodes

Abstract

Background: The lymphatic system is commonly injured during cancer treatment. However, despite the morbidity of these injuries, there are currently no options for replacing damaged lymphatics. The purpose of this study was to optimize methods for decellularization of murine lymph nodes (LN) and to determine if these scaffolds can be used to tissue engineer lymph node-like structures. Methods and Results: LNs were harvested from adult mice and subjected to various decellularization protocols. The degree of decellularization and removal of nuclear material was analyzed histologically and quantitatively using DNA isolation. In addition, we analyzed histological architecture by staining for matrix proteins. After the optimal method of decellularization was identified, decellularized constructs were implanted in the renal capsule of syngeneic or allogeneic recipient mice and analyzed for antigenicity. Finally, to determine if decellularized constructs could deliver lymphocytes to recipient animals, the matrices were repopulated with splenocytes, implanted in submuscular pockets, and harvested 14 days later. Decellularization was best accomplished with the detergent sodium dodecyl sulfate (SDS), resulting in negligible residual cellular material but maintenance of LN architecture. Implantation of decellularized LNs into syngeneic or allogeneic mice did not elicit a significant antigenic response. In addition, repopulation of decellularized LNs with splenocytes resulted in successful in vivo cellular delivery. Conclusions: We show, for the first time, that LNs can be successfully decellularized and that these matrices have preserved extracellular matrix architecture and the potential to deliver leukocytes in vivo. Future studies are needed to determine if tissue engineered lymph nodes maintain immunologic function.

Decellularized Lymph Node Scaffolding as a Carrier for Dendritic Cells to Induce Anti-Tumor Immunity

“…The utility of decellularized scaffolds has widely focused on the tendon, bladder, blood vessels, kidney, and lung, but few on the lymph node [30,31,32]. Cuzzone et al demonstrated that lymph nodes could be decellularized by sodium dodecyl sulfate treatment, successfully eliminating the cellular material and preserving the ECM [16]. However, other common decellularized methods, such as TRITON-X and 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate, were not effective when used in lymph nodes.…”

“…It provides a strategy by which to fabricate lymph node scaffolds on which to grow immune cells with immunological function. Previous studies demonstrated that spleen-derived decellularized ECM or lymph node-associated decellularized ECM can deliver immune cells and promote the organization of lymphoid-like scaffolds [16,17]; however, whether these can induce antigen-specific immunity or the antitumor immune response is still unknown.…”

Deconstructing Immune Microenvironments of Lymphoid Tissues for Reverse Engineering

“…The environment itself, the stroma, seems to be crucial. Implanted scaffolds with lymphoid tissue initiators, or implanting repopulated decellularized lymphoid organs, have presented some successful results in animal models. Engineering lymphoid stroma has also been pursued in vitro .…”

In vivo construction of lymphoid node by implantation of adipose-derived stromal cells with hydroxypropyl methyl cellulose hydrogel in BALB/c nude mice

“…Daniel et al engineered lymph nodes by using SDS to decellularize the lymph node of mice and repopulated it with leukocytes harvested from the spleen of another donor mouse. 2 Purwada et al first engineered gelatin hydrogels combined with polyionic silicate nanoparticles to encapsulate murine germinal center B cell clusters in 2D cultures. 3 However, there are no widely accepted techniques at present.…”