Removal of minute virus of mice-mock virus particles by nanofiltration of culture growth medium supplemented with 10% human platelet lysate |
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| Institution: | 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 Program in Cell Therapy and Regeneration Medicine, Taipei Medical University, Taipei, Taiwan;1. Laboratory of Biochemistry and Immunology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea;2. Laboratory Animal Research Center, Chungbuk National University, Cheongju, Republic of Korea;3. R&D Center, Autotelic Bio, Inc, Seongnam, Republic of Korea;4. Laboratory of Pharmacology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Republic of Korea;1. Anthony Nolan Cell Therapy Centre, Nottingham, UK;2. The Anthony Nolan Research Institute, Nottingham, UK;1. National Center for Cancer Immune Therapy, Department of Oncology, Copenhagen University Hospital, Herlev, Denmark;2. Department of Hematology, Rigshospitalet, Copenhagen, Denmark;1. Department of Regenerative Technologies and Biofabrication, National Medical Research Radiological Center, Obninsk, Russia;2. Basel University, Basel, Switzerland;1. Department of Pediatrics, Division of Blood and Marrow Transplantation, University of Minnesota Cancer Center, Minneapolis, Minnesota, USA;2. Ben Towne Center for Childhood Cancer Research, Seattle Children''s Research Institute, Seattle, Washington, USA;3. Department of Pediatrics, University of Washington, Seattle, Washington, USA;4. Laboratory of Lymphocyte Signaling and Development, Babraham Institute, Cambridge, UK;5. Translational Oncology, Allogene Therapeutics, San Francisco, California, USA;6. Department of Medicine, Division of Hematology/Oncology and Transplantation, University of Minnesota, Minneapolis, Minnesota, USA;7. Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA;8. Division of Pediatric Hematology/Oncology, Boston Children''s Hospital and Dana-Farber Cancer Institute, Boston, Massachusetts, USA |
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| Abstract: | Background aimsPlatelet 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.MethodsHerein, 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.ResultsNo 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.ConclusionsThe 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. |
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