A 3D bioprinter platform for mechanistic analysis of tumoroids and chimeric mammary organoids

Abstract

The normal mammary microenvironment can suppress tumorigenesis and redirect cancer cells to adopt a normal mammary epithelial cell fate in vivo . Understanding of this phenomenon offers great promise for novel treatment and detection strategies in cancer, but current model systems make mechanistic insights into the process difficult. We have recently described a low-cost bioprinting platform designed to be accessible for basic cell biology laboratories. Here we report the use of this system for the study of tumorigenesis and microenvironmental redirection of breast cancer cells. We show our bioprinter significantly increases tumoroid formation in 3D collagen gels and allows for precise generation of tumoroid arrays. We also demonstrate that we can mimic published in vivo findings by co-printing cancer cells along with normal mammary epithelial cells to generate chimeric organoids. These chimeric organoids contain cancer cells that take part in normal luminal formation. Furthermore, we show for the first time that cancer cells within chimeric structures have a significant increase in 5-hydroxymethylcytosine levels as compared to bioprinted tumoroids. These results demonstrate the capacity of our 3D bioprinting platform to study tumorigenesis and microenvironmental control of breast cancer and highlight a novel mechanistic insight into the process of microenvironmental control of cancer.

3D bioprinting for reconstituting the cancer microenvironment

“… Cancer model type Cell types used Bioink or substrate used Bioprinting modalities used Ref. Glioblastoma-on-a-chip Glioma cell line U118 and endothelial cells Collagen or dECM hydrogel EBB 28 Glioma stem cell (shell); glioma cell line (core) Alginate Coaxial EBB 29 Glioma stem cells Gelatin, alginate, and fibrinogen EBB 30 Hepatoma HepG2 and glioma cell U251 Alginate DBB (Inkjet) 31 iPSC-derived human neural progenitor cells and U118 human glioma cells Scaffold-free 3D culture EBB 32 Human glioma stem cell line, U118 Sodium alginate and gelatin EBB 33 Glioblastoma-associated macrophages (GAMs) and glioblastoma cells GelMA EBB 34 Human primary umbilical cord-derived mesenchymal stromal cells (UC-MSC, referred to as MSCs), HUVEC, and human bone marrow-derived epithelial-neuroblastoma immortalized cells (SH-SY5Y) Agarose and type-I collagen DBB 100 Breast tumor model Immortalized non-tumorigenic human breast epithelial cell lines MCF-12A and MCF10A Rat tail collagen I EBB 36 MCF-7 BC cells PBS solution DBB 101 Immortalized non-tumorigenic human breast epithelial cell line, MCF-12A, and the breast carcinoma cell lines MCF-7 and MDA-MB-468 Rat tail collagen EBB 37 MCF-7 cell Gelatin-PEG DBB 38 BT474 breast cancer cells, human perinatal foreskin fibroblasts (BJ), and human adult dermal fibroblasts (HDF) Poly(ethylene glycol) diacrylate (PEGDA) LBB (optical project...…”

Cell Culture Based in vitro Test Systems for Anticancer Drug Screening

“…It is known that 2D cultures do not fully reflect the pathophysiology of tumor cells and the actual level of resistance (Shoemaker, 2006) JFCR39 panel (Nakatsu et al, 2007) Boyden's chamber Possibility to study the effect of the test substance on the invasiveness and migration potential of tumor cells (Reid et al, 2019) to radiotherapy or chemotherapy in the tumor niche in the in vivo system (Chen et al, 2012; Table 1). Studies have shown that gene expression profiles as well as treatment responses in multicellular spheroid 3D models are more similar to the in vivo situation (Riedl et al, 2017).…”

Mimicking tumor hypoxia and tumor-immune interactions employing three-dimensional in vitro models

“…To address these issues, 3D cell culture models are a reliable alternative, providing experimentally accessible human models to study the biological processes of cancer. Several 3D culture platforms such as spheroids, organoids, hydrogels, 3D scaffolds, 3D bio-printing, and microfluidics have attempted the recreation of certain aspects of tumor microenvironments present in tissues including the brain [30][31][32][33][34][35][36], breast [37][38][39][40][41][42][43], ovarian [44][45][46][47][48][49][50][51], bone [52][53][54][55][56][57][58], liver [59][60][61][62][63][64][65], lung [66][67][68][69][70][71][72], colon [73][74][75][76]…”