Production of LN-521 coated microcarriers for stem cell expansion

Purpose and goal

The project aimed at enabling a standardized and scalable stem cell culture for cell therapy, drug development and toxicology studies. We will develop and commercialize LN-521 coated hydrogel microcarriers and protocols for the expansion of human pluripotent stem cells (hPSC) in bioreactors. The main goals were to develop: 1. A method to scale up the production of recombinant LN-521 2. A hydrogel microcarrier coated with LN-521 adapted to support hPSC growth 3. An optimized protocol to grow hPSCs in bioreactors

Expected results and effects

The project partners were to first develop a GMP LN-521 coating substrate. Thereafter, develop a new hydrogel-based microcarrier coated with LN-521 for scalable hPSC culture in bioreactors. Both GMP LN521 and the LN-521-coated hydrogel beads were going to be commercialized. We have managed to express LN-521 in fully documented and thus sufficiently traceable production cells. Currently we validate protein function and have scheduled a GMP pilot run at a certified GMP CMO. Furthermore, we have identified a hydrogel that supports stem cell expansion when covalently linked to LN-521.

Planned approach and implementation

The project is divided into 4 WPs: 1. Scale up LN-521 production Methods: A suspension-based cell line called CAP has been used Analysis: The CAP cells produces a functional protein in large scale 2. Development of a microcarrier coated with LN-521 Methods: Screen different hydrogels, make beads and culture hPSCs in bioreactor Analysis: Optimal material found. Bead manufacturing hard. Another product in pipe 3. Optimization of hPSC bioreactor culture conditions Methods: Set specifications; static small scale first Analysis: Optimal protocol set 4. Pre-production Not initiated