Drug Discovery 2022: driving the next life science revolution
Poster
135

New organoid scaffold with fibrillar collagen, laminin, and hyaluronic acid

Authors

K Mizuno1; Y Murasawa1; K Fujita1; Y Kumazawa1; T Sasaki2; Y Takemura3; A Sim4
1 Nippi Research Institute of Biomatrix, Japan;  2 Oita University, Japan;  3 KyoDiagnosis, Japan;  4 AMSBIO, UK

Abstract

3D culture is essential for mimicking real tissues and organs. At present, mouse basement membrane extract (BME/Matrigel) is the standard substrate used in vitro. Major components are laminin-111, type IV collagen-121, nidogen-1 and HSPG. 



To develop a new substrate that mimics the in vivo environment, we searched for a solution which is liquid at low temperatures and gels at the temperature of the cell culture.

Our gel is based on fibrillar collagen and the E8 fragment of laminin cross-linked to hyaluronic acid with genipin.



The effects of genipin concentration, molecular weight and concentration of hyaluronic acid, and the type (acid soluble/pepsin-solubilized) and concentration of collagen were examined.



In addition to the basic formula, collagen (acid soluble, pepsin extracted; I&III, IV, V) and laminin-E8 fragment isoforms (111, 332, & 511) were used to adjust interaction with cell-specific receptors.



When 12-day mouse embryo cells isolated from kidney, colon, and liver were cultured in the gel, they differentiated and organized.

Mouse embryonic-derived renal cells cultured in the gel were monodisperse immediately after culture, but aggregation was observed after one day, and self-assembly into tissues with branching ureter was advanced by Day 7. Staining, with a marker specific for ureteral tissue, showed the branched structure.



The concept is to build physiological connective-tissue ECM including basement membrane and interstitial collagen fibrils.



This new gel can be used as an alternative to Matrigel/BME, overcoming some of the shortcomings of BME. It has more versatile properties that can regulate both mesenchymal and epithelial tissues in the body. 



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