Expansion of human mesenchymal stem cells on poly(vinyl alcohol) microcarriers

Masahiro Kaneko; Airi Sato; Satoru Ayano; Akio Fujita; Goro Kobayashi; Akira Ito

doi:10.1016/j.jbiosc.2023.08.003

Expansion of human mesenchymal stem cells on poly(vinyl alcohol) microcarriers

J Biosci Bioeng. 2023 Nov;136(5):407-414. doi: 10.1016/j.jbiosc.2023.08.003. Epub 2023 Aug 30.

Authors

Masahiro Kaneko¹, Airi Sato², Satoru Ayano³, Akio Fujita³, Goro Kobayashi³, Akira Ito⁴

Affiliations

¹ Department of Chemical Systems Engineering, School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
² College of Bioscience and Biotechnology, Chubu University, 1200 Matsumoto, Kasugai, Aichi 487-8501, Japan.
³ Research and Development Division, Kuraray Co., Ltd., 41 Miyukigaoka, Tsukuba, Ibaraki 305-0841, Japan.
⁴ Department of Chemical Systems Engineering, School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan. Electronic address: ito.akira@material.nagoya-u.ac.jp.

PMID: 37657971
DOI: 10.1016/j.jbiosc.2023.08.003

Abstract

Microcarriers provide a high surface-area-to-volume ratio that can realize high yields of cell products, including human mesenchymal stem cells (hMSCs). Here, we report a novel poly(vinyl alcohol) (PVA)-based microcarrier for hMSC expansion in suspension culture. PVA microcarriers were prepared as collagen-coated PVA hydrogels 181 μm in size and a high surface-area-to-weight ratio of 2945 cm²/g. The PVA microcarriers supported a 2.6-fold expansion of hMSCs in a 30-mL single-use stirred bioreactor after a 7 d culture period, comparable to that of commercially available microcarriers. Interestingly, we observed that hMSCs on PVA microcarriers adhered to adjacent microcarriers, resulting in the aggregation of hMSC-PVA microcarriers. Therefore, we conducted a long-term expansion culture using a bead-to-bead cell transfer method with PVA microcarriers. Fresh microcarriers were added to the cell-populated microcarriers in the bioreactor on days 7 and 14. hMSCs on PVA microcarriers continued to grow for 21 d using the bead-to-bead cell transfer method. Furthermore, magnetic PVA (PVA-mag) microcarriers were developed by loading magnetic nanoparticles into PVA microcarriers, and we demonstrated that these PVA-mag microcarriers enabled cell recovery by magnetic separation. These results suggest that these PVA microcarriers can contribute to the large-scale culture of hMSCs for regenerative medicine and cell therapy.

Keywords: Bioreactor; Magnetic nanoparticle; Magnetic separation; Mesenchymal stem cell; Microcarrier; Poly(vinyl alcohol).