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Nanofiltration of growth media supplemented with human platelet lysates for pathogen-safe xeno-free expansion of mesenchymal stromal cells.
Barro, Lassina; Nebie, Ouada; Chen, Ming-Sheng; Wu, Yu-Wen; Koh, Mickey Bc; Knutson, Folke; Watanabe, Naoto; Takahara, Masayasu; Burnouf, Thierry.
  • Barro L; International PhD Program in Biomedical Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan.
  • Nebie O; Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan.
  • Chen MS; Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan.
  • Wu YW; Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan.
  • Koh MB; Department of Haematology, St George's University Hospitals Foundation NHS Trust, London, UK; Blood Sciences Group, Health Sciences Authority, Singapore.
  • Knutson F; Clinical Immunology and Transfusion Medicine IGP, Uppsala University, Uppsala, Sweden.
  • Watanabe N; Asahi Kasei Medical Co., Ltd., Tokyo, Japan.
  • Takahara M; Asahi Kasei Medical Co., Ltd., Tokyo, Japan.
  • Burnouf T; 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
Cytotherapy ; 22(8): 458-472, 2020 08.
Article in English | MEDLINE | ID: covidwho-209852
Semantic information from SemMedBD (by NLM)
1. Plasma LOCATION_OF Platelet concentrate
Subject
Plasma
Predicate
LOCATION_OF
Object
Platelet concentrate
2. Extracellular Vesicles PART_OF Blood Platelets
Subject
Extracellular Vesicles
Predicate
PART_OF
Object
Blood Platelets
3. Plasma LOCATION_OF Platelet concentrate
Subject
Plasma
Predicate
LOCATION_OF
Object
Platelet concentrate
4. Extracellular Vesicles PART_OF Blood Platelets
Subject
Extracellular Vesicles
Predicate
PART_OF
Object
Blood Platelets
ABSTRACT
BACKGROUND

AIMS:

Human platelet lysate can replace fetal bovine serum (FBS) for xeno-free ex vivo expansion of mesenchymal stromal cells (MSCs), but pooling of platelet concentrates (PCs) increases risks of pathogen transmission. We evaluated the feasibility of performing nanofiltration of platelet lysates and determined the impact on expansion of bone marrow-derived MSCs.

METHODS:

Platelet lysates were prepared by freeze-thawing of pathogen-reduced (Intercept) PCs suspended in 65% storage solution (SPP+) and 35% plasma, and by serum-conversion of PCs suspended in 100% plasma. Lysates were added to the MSC growth media at 10% (v/v), filtered and subjected to cascade nanofiltration on 35- and 19-nm Planova filters. Media supplemented with 10% starting platelet lysates or FBS were used as the controls. Impacts of nanofiltration on the growth media composition, removal of platelet extracellular vesicles (PEVs) and MSC expansion were evaluated.

RESULTS:

Nanofiltration did not detrimentally affect contents of total protein and growth factors or the biochemical composition. The clearance factor of PEVs was >3 log values. Expansion, proliferation, membrane markers, differentiation potential and immunosuppressive properties of cells in nanofiltered media were consistently better than those expanded in FBS-supplemented media. Compared with FBS, chondrogenesis and osteogenesis genes were expressed more in nanofiltered media, and there were fewer senescent cells over six passages.

CONCLUSIONS:

Nanofiltration of growth media supplemented with two types of platelet lysates, including one prepared from pathogen-reduced PCs, is technically feasible. These data support the possibility of developing pathogen-reduced xeno-free growth media for clinical-grade propagation of human cells.
Subject(s)
Keywords

Full text: Available Collection: International databases Database: MEDLINE Main subject: Blood Platelets / Cell Culture Techniques / Nanotechnology / Mesenchymal Stem Cells / Filtration Limits: Humans Language: English Journal: Cytotherapy Journal subject: Therapeutics Year: 2020 Document Type: Article Affiliation country: J.jcyt.2020.04.099

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Blood Platelets / Cell Culture Techniques / Nanotechnology / Mesenchymal Stem Cells / Filtration Limits: Humans Language: English Journal: Cytotherapy Journal subject: Therapeutics Year: 2020 Document Type: Article Affiliation country: J.jcyt.2020.04.099