Abstract

Controlled Development of Functional Cerebellar Organoids on Synthetic Peptide-based Hydrogels

Aiste Jekabsone, Researcher, Lithuanian University of Health Sciences

There are few ECM-mimicking cell culture options that would be as easy to use and analytically efficient as monolayers on glass or plastic. Aiming to meet this demand, shape-retaining and transparent collagen mimetic peptide-polyethylene glycol based hydrogels were applied as multiwell plate inserts for in vitro cell culture. The hydrogels promoted development of cerebellar explants to spheroidal bodies resembling cerebellar granule layer organization with incorporated velate astrocytes and mobile round-shaped or settled ramified microglia. Neurons in the spheroids formed functional networks observed as Ca²⁺ oscillation with frequency similar to the in vivo activity in cerebellum. Cell attachment, organization, neuritogenesis and neurite branching were modulated by altering the polymer concentration in hydrogel and introducing extracellular matrix peptides RGD and IKVAV. Microformation was used to control spatial organization and size of the spheroidal bodies. The study revealed that matrices developed are easy to operate and compatible with most types of microscopy and common plate readers. These properties allow for easy switching from plastic to more in vivo-relevant environment while retaining convenience and analytical capacity. The functionalization by bioactive peptides and microformation suggests these matrices as a tool for ECM research, tissue engineering and regenerative medicine.