Microparticles for Suspension Culture of Mammalian Cells

Abstract

The focus of this work is to develop a technology for the synthesis of polymer microcarriers that demonstrate mammalian cell culture adhesion on the surface of the microcarriers. Most mammalian cells are adherent in nature that requires multilayer vessels, large volume, expensive cell culture media, high manufacturing time, and high costs of cell culture supplies for the commercial-scale manufacturing of cells. The development of an efficient, scalable technology for producing large volumes of cells is a need in bioprocess industries to improve product potency. We developed a method of synthesizing soft biocompatible US FDA approved polymer based microparticle carrier system of approximately 260 ± 27 μm in diameter that serves as an adherent platform for human umbilical vein endothelial cells (HUVEC) to grow in suspension. Our preliminary experimental results showed that using the polymeric microcarrier system cell adhesion to the surface of the microcarriers was 2-3-fold higher than conventional cell culture flasks while using 10-fold lower cell culture media in a bioreactor than a tissue-culture treated flask. The survival of HUVEC on microparticles was confirmed by live cell staining (green fluorescent calcein AM), dead cell staining (ethidium homodimer-1), nuclear DAPI staining, actin cytoskeleton staining, confocal microscopy, and flow cytometry analysis. This technology will provide high cell culture productivity while reducing the costs of growing adherent cells.