Removal process of prion and parvovirus from human platelet lysates used as clinical-grade supplement for ex vivo cell expansion

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• 作者：Yu-Chun Kao¹ ; Andy Bailey² ; Bernhard Samminger² ; Junji Tanimoto³ ; Thierry Burnouf¹ ; ^{thburnouf@gmail.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：cell therapy ; chromatography ; hollow fiber ; membrane ; platelet lysate ; prion ; safety ; virus
• 刊名：Cytotherapy
• 出版年：2016
• 出版时间：July 2016
• 年：2016
• 卷：18
• 期：7
• 页码：911-924
• 全文大小：4710 K

文摘

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.

Methods

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.

Results

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.

Conclusions

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.