In vitro ovarian cancer model based on three-dimensional agarose hydrogel

Abstract

To establish a typical tumor model of ovarian cancer which may be more representative and reliable than traditional monolayer culture and pellet, agarose was used as cell vehicle to engineering tumor. Selection of agarose is based on its successful application in tissue engineering with both amenable mechanical and biological properties. In this study, ovarian cancer cell line SKOV3 was encapsulated in agarose hydrogel with cell aggregates and two-dimensional culture as controls. In vitro cell proliferation was assessed by MTT and cell viability was examined at time points of 2, 4, and 6 days. The expression of tumor malignancy markers including matrix metalloproteinase 2, matrix metalloproteinase 9, hypoxia-inducible factor-1α, and vascular endothelial growth factor–A was assessed by real-time polymerase chain reaction. The results showed that cells proliferated more rapidly in three-dimensional agarose culture than controls. Furthermore, upregulation of matrix metalloproteinase 9 and matrix metalloproteinase 2 activity and increased expression of vascular endothelial growth factor–A and hypoxia-inducible factor-1α were shown in agarose-engineered tumors. All the evidences demonstrated that agarose may provide a more favorable environment for cancer cell growth, mimicking the in vivo environment for tumor generation. The novel in vitro tumor model may be useful for the further investigation of anticancer therapeutics.

Modelling the helium plasma jet delivery of reactive species into a 3D cancer tumour

“…By introducing fluid flow into the experiment, perhaps with a small pumping system, it would be interesting to see how far RONS species persist from the He plasma jet treatment site. Second, agarose is devoid in biological material, but it is possible to add biomolecules to mimic different tissues [125], or to incorporate live cells into the agarose matrix to mimic 3D tumours [126,127].…”

Porcine spermatogonial stem cells self‐renew effectively in a three dimensional culture microenvironment

“…However, agarose‐based 3D hydrogels are more difficult to manipulate at physiological temperatures than at higher temperatures (Aymard et al, ), which can lead to untimely coagulation of the agarose (Elhija et al, ). There are also difficulties with retrieving all of the cultured cells from the hydrogel (Elhija et al, ; Xu et al, ) and incorporating ECM analogs (Mercey et al, ), making long‐term culture difficult. Thus, further research using biodegradable and biocompatible materials, which can overcome the limitations of agarose‐based 3D hydrogels, is required.…”

Recreating complex pathophysiologies in vitro with extracellular matrix surrogates for anticancer therapeutics screening

“…For example, hyaluronic acid (HA) hydrogels loaded with LNCaP (prostate) cells have been shown to stimulate development of multicellular tumoroids for the assessment of the effectiveness of doxorubicin-loaded nanoparticles and to simulate tissue metastasis at an early stage [144,145]. Agarose hydrogels have been used with SKOV3 (ovarian) cancer cell aggregates and demonstrated increased cell proliferation, compared with 2D controls, and upregulated tumour malignancy markers (e.g., MMP2, MMP9, hypoxiainducible factor (HIF)-1α, vascular endothelial growth factor (VEGF)-α] [146]. Although chemoresistance of collagen hydrogels loaded with HT29 (colorectal), CF56 (colon), human umbilical vein endothelial and 3T3 cells was not assessed for colorectal tumour, dense tumour-like masses were created upon application of plastic compression that increased cell and matrix density [147].…”