Decellularization of xenografted tumors provides cell-specific in vitro 3D environment

Abstract

In vitro cell culture studies are common in the cancer research field, and reliable biomimetic 3D models are needed to ensure physiological relevance. In this manuscript, we hypothesized that decellularized xenograft tumors can serve as an optimal 3D substrate to generate a top-down approach for in vitro tumor modeling. Multiple tumor cell lines were xenografted and the formed solid tumors were recovered for their decellularization by several techniques and further characterization by histology and proteomics techniques. Selected decellularized tumor xenograft samples were seeded with the HCC1806 human triple-negative breast cancer (TNBC) basal-like subtype cell line, and cell behavior was compared among them and with other control 2D and 3D cell culture methods. A soft treatment using Freeze-EDTA-DNAse allows proper decellularization of xenografted tumor samples. Interestingly, proteomic data show that samples decellularized from TNBC basal-like subtype xenograft models had different extracellular matrix (ECM) compositions compared to the rest of the xenograft tumors tested. The in vitro recellularization of decellularized ECM (dECM) yields tumor-type–specific cell behavior in the TNBC context. Data show that dECM derived from xenograft tumors is a feasible substrate for reseeding purposes, thereby promoting tumor-type–specific cell behavior. These data serve as a proof-of-concept for further potential generation of patient-specific in vitro research models.

Decellularized diseased tissues: current state‐of‐the‐art and future directions

“…Additionally, it preferably displays the structure of tumor and the complexity of TME 66 . Decellularized tumor ECM‐based 3D tumor models have been shown to be a promising platform for exploring the growth of tumor cells in vitro, the release of supporting growth factors and the screening of related tumor genes 38,67,68 . In the process of applying decellularized ECM to 3D tumor model, maintaining long‐term cell activity is crucial, and there are many factors affecting the results, such as different culture conditions (pH value, matrix mechanical properties, temperature, etc.)…”