Removal of minute virus of mice-mock virus particles by nanofiltration of culture growth medium supplemented with 10% human platelet lysate

Lassina Barro; Liling Delila; Ouada Nebie; Yu-Wen Wu; Folke Knutson; Naoto Watanabe; Masayasu Takahara; Thierry Burnouf

doi:10.1016/j.jcyt.2021.05.006

Removal of minute virus of mice-mock virus particles by nanofiltration of culture growth medium supplemented with 10% human platelet lysate

Cytotherapy. 2021 Oct;23(10):902-907. doi: 10.1016/j.jcyt.2021.05.006. Epub 2021 Jul 5.

Authors

Lassina Barro¹, Liling Delila², Ouada Nebie², Yu-Wen Wu², Folke Knutson³, Naoto Watanabe⁴, Masayasu Takahara⁴, Thierry Burnouf⁵

Affiliations

¹ International PhD Program in Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan.
² Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan.
³ Clinical Immunology and Transfusion Medicine IGP, Uppsala University, Uppsala, Sweden.
⁴ Asahi Kasei Medical Co, Ltd, Tokyo, Japan.
⁵ International PhD Program in Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan; Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan; International Program in Cell Therapy and Regeneration Medicine, Taipei Medical University, Taipei, Taiwan. Electronic address: thburnouf@gmail.com.

PMID: 34238658
DOI: 10.1016/j.jcyt.2021.05.006

Abstract

Background aims: Platelet concentrates (PCs) are pooled to prepare human platelet lysate (HPL) supplements of growth media to expand primary human cells for transplantation; this increases the risk of contamination by known, emerging, and unknown viruses. This possibility should be of concern because viral contamination of cell cultures is difficult to detect and may have detrimental consequences for recipients of cell therapies. Viral reduction treatments of chemically defined growth media have been proposed, but they are not applicable when media contain protein supplements currently needed to expand primary cell cultures. Recently, we successfully developed a Planova 35NPlanova 20N nanofiltration sequence of growth media supplemented with two types of HPL. The nanofiltered medium was found to be suitable for mesenchymal Stromal cell (MSC) expansion.

Methods: Herein, we report viral clearance achieved by this nanofiltration process used for assessing a new experimental model using non-infectious minute virus of mice-mock virus particle (MVM-MVP) and its quantification by an immunoqPCR. Then, high doses of MVM-MVP (1012 MVPs/mL) were spiked to obtain a final concentration of 1010 MVPs/mL in Planova 35N-nanofiltered growth medium supplemented with both types of HPLs [serum converted platelet lysate SCPL) and intercept human platelet lysate (I-HPL)] at 10% (v/v) and then filtering through Planova 20N.

Results: No substantial interference of growth medium matrices by the immune-qPCR assay was first verified. Log reduction values (LRVs) were ≥ 5.43 and ≥ 5.36 respectively, SCPL and I-HPL media. MVM-MVPs were also undetectable by dynamic light scattering and transmission electron microscopy.

Conclusions: The nanofiltration of growth media supplemented with 10% HPL provides robust removal of small nonenveloped viruses, and is an option to improve the safety of therapeutic cells expanded using HPL supplements.

Keywords: HPL; human platelet lysate; log reduction value; nanofiltration; viral clearance.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Culture Techniques
Culture Media
Humans
Mesenchymal Stem Cells*
Mice
Minute Virus of Mice*
Virion

Substances

Culture Media