The transplanted lymph node was harvested from mice and its functionality was evaluated ex-vivo.
Characterization of Immune Cell Population
In flow cytometry, T cells were selected by CD3 marker, and myeloid cells were selected by CD11b marker (A). The distributions of T cells and myeloid cells were similar between transplanted decellularized lymph nodes and lymph nodes with sham surgery, as the histograms were overlapped (B). Quantification of T cells (for both CD4 and CD8 subsets) and myeloid cells also showed no significant difference between transplanted decellularized lymph nodes and lymph nodes with sham surgery (C). This result validates that our lymph nodes homed similar cell populations as a native lymph node.
Vascularization Characterization
Vasculature formation could be observed around and inside the lymph nodes (Green).
The amount of vasculature formation was quantified by measuring the area stained green divided by the total area of the lymph node. Quantification showed no significant difference between transplanted decellularized lymph nodes and lymph nodes with sham surgery.
The lymph node was able to implement into nearby vasculature and home immune cells to restore immune functions