Functional activity of insulinoma cells (INS‐1E) and pancreatic islets cultured in agarose cryogel sponges

Abstract

Here, we describe the preparation, structure, and properties of cryogel sponges, which represent a new type of macroporous biomaterial for tissue engineering. Cryogels were produced through freeze-thawing techniques, either from agarose alone or from agarose with grafted gelatin. The aim of this study was to evaluate agarose cryogel sponges as scaffolds for culturing both isolated pancreatic islets and insulinoma cells (INS-1E). In order to evaluate the effect of cell entrapment in artificial scaffolds, cell function reflected by insulin secretion and content was studied in cells cultivated for a 2-week period either in culture plastic plates or in cryogel sponge disks. Our results show that tumor-derived INS-1E cells grown either on plastic or on cryogels do not differ in their proliferation, morphology, insulin release, and intracellular insulin content. However, isolated pancreatic islets cultivated on cryogels sponge show 15-fold higher basal insulin secretion at 3.0 mM glucose than islets cultivated on plastic plates and fail to respond to stimulation with 16.7 mM glucose. In addition, these islets have about 2-fold lower insulin content compared to those grown in plastic plates. It is possible that the cell dysfunction noted in these in vitro experiments is due to the effect of the limited oxygen supply to the islets cultivated in cryogel sponge. Further in vivo studies are needed to clarify the nature of such an observation since according to previous reports, agarose and gelatin induce new vessel formation supporting enhanced oxygen supply.

Cryostructuring of polymer systems. XXIX. Preparation and characterization of supermacroporous (spongy) agarose‐based cryogels used as three‐dimensional scaffolds for culturing insulin‐producing cell aggregates

“…In this study, we showed how some of these factors can influence the properties and morphology of ABCGs. In addition, the data of our previous work, 7 as well as the results of ABCG biotesting carried out in this study, offer the promise of potential applications of these supermacroporous matrices as biocompatible 3D scaffolds for cell culture. A. V. Kaznacheev (A. N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences) for help with the microscopy experiments.…”

Cytocompatibility studies of mouse pancreatic islets on gelatin - PVP semi IPN scaffolds in vitro: Potential implication towards pancreatic tissue engineering

“…In the There were reports of growing islets for 49 days on chitoson sponge, 29 and 2 weeks on agarose cryogel. 30 So the behavior of mouse islets on gelatin/PVP scaffolds were determined in vitro. The direct contact assay showed that the cells were viable on all the scaffolds up to 48 h. In order to view the in growth of cells towards interior portion of the scaffolds, z sectioning was done present scenario, it is expected that the presence of PVP is an added benefit for maintaining structure and function of islet cells.…”

Macroporous gels prepared at subzero temperatures as novel materials for chromatography of particulate‐containing fluids and cell culture applications

“…Their main applications are found in biocatalysis with immobilized cells [12,27,54,64,65] and enzymes [38,57,58], in bioseparations for purification of targets from particulatecontaining fluids [18, 30 -33, 37, 66, 67], chromatography of cell organelles [35], viruses [68], microbial [28,61,62], and mammalian [16,19,40] cells and in biomedical applications as 3-D matrixes for culture of mammalian cells [41,69].…”