Removal process of prion and parvovirus from human platelet lysates used as clinical-grade supplement for ex vivo cell expansion.
Pooled human platelet lysate (HPL) is becoming the new gold standard as supplement for ex vivo cell culture for clinical protocols. However, the risk of pathogen contamination of HPL increases with the platelet pool size. We hypothesized that hollow fiber anion exchange membrane chromatography using QyuSpeed D (QSD) could remove resistant and untested bloodborne pathogens, such as parvoviruses and prions, from HPL-supplemented growth media without substantially affecting their capacity to support ex vivo cell expansion.



Frozen or thawed platelet concentrates were serum-converted and centrifuged for obtaining HPL that was added to various growth media (ca. 100 mL), filtered through a 0.6-mL QSD membrane and characterized for proteins, growth factors and chemical composition. Capacity to expand Chinese hamster ovary, periodontal ligament, gingival fibroblast cells and Wharton's jelly mesenchymal stromal cells was studied. Removal of porcine parvovirus (PPV) and of the 263K prion strain of hamster-adapted scrapie was studied by spiking experiments following international guidelines.



QSD had minimal impact on HPL-supplemented medium composition in proteins, growth factors and chemical content, nor capacity to expand and differentiate cells. In addition, QSD could remove ≥5.58 log10 [TCID50/mL] and ≥3.72 log10 of PPV and the 263K prion, respectively.



QSD hollow fiber chromatography can be used to improve the virus and prion safety of HPL-supplemented media to safely expand cells for clinical protocols. These data bring new perspectives for increasingly safer use of pooled HPL in cell therapy and regenerative medicine applications.
Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan.
Yu-Chun
YC
Kao
ViruSure, Tech Gate Science and Technology Park, Donau City Strasse 1, A-1220, Vienna, Austria.
Andy
A
Bailey
ViruSure, Tech Gate Science and Technology Park, Donau City Strasse 1, A-1220, Vienna, Austria.
Bernhard
B
Samminger
Asahi Kasei Medical Co, Ltd., Tokyo, Japan.
Junji
J
Tanimoto
Graduate Institute of Biomedical Materials and Tissue Engineering, College of Biomedical Engineering, Taipei Medical University, Taipei, Taiwan. Electronic address: thburnouf@gmail.com.
Thierry
T
Burnouf
D000818:Animals; D001792:Blood Platelets / Q000166:cytology* / Q000821:virology*; D016466:CHO Cells; D002452:Cell Count; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002470:Cell Survival; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media; D005347:Fibroblasts / Q000166:cytology; D005881:Gingiva / Q000166:cytology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology; D017992:Parvovirus / Q000302:isolation & purification*; D010513:Periodontal Ligament / Q000166:cytology; D011328:Prions / Q000302:isolation & purification*; D034421:Sus scrofa; D059631:Wharton Jelly / Q000166:cytology
D003470:Culture Media; D011328:Prions
cell therapy; chromatography; hollow fiber; membrane; platelet lysate; prion; safety; virus