Pathogen reduction through additive-free short-wave UV light irradiation retains the optimal efficacy of human platelet lysate for the expansion of human bone marrow mesenchymal stem cells

Viau, Sabrina; Chabrand, Lucie; Eap, Sandy; Lorant, Judith; Rouger, Karl; Goudaliez, F.; Sumian, Chryslain; Delorme, Bruno

doi:10.1371/journal.pone.0181406

Cited by 30 publications

(33 citation statements)

References 72 publications

(99 reference statements)

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“…Moreover, the interaction of BM‐MSCs versus PBMNCs showed an inhibition of PBMNC proliferation. This inhibition was described in previous studies at the 1:20 ratio but is similar to ours at the 1:10 ratio. This inhibition was less with DVI‐HPL.…”

Section: Discussionsupporting

confidence: 92%

A double‐virally‐inactivated (Intercept–solvent/detergent) human platelet lysate for in vitro expansion of human mesenchymal stromal cells

Barro

Nebie

et al. 2019

Transfusion

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BACKGROUND: Pooled human platelet lysate (HPL) can replace fetal bovine serum (FBS) as xeno-free supplement for ex vivo expansion of mesenchymal stromal cells (MSCs). We evaluate here whether a double-virallyinactivated HPL (DVI-HPL) prepared from expired Intercepttreated platelet concentrates (PCs) and treated by solvent/detergent (S/D) can be used for MSC expansion.STUDY DESIGN AND METHODS: Expired Intercepttreated PCs in 65% platelet (PLT) additive solution were pooled and subjected to a 1% tri-n-butyl phosphate/1% Triton X-45 treatment followed by soybean oil, hydrophobic interaction chromatography purification, and sterile filtration. Bone marrow-derived MSCs (BM-MSCs) were expanded for four passages in growth medium containing 10% DVI-HPL, I-HPL (from Intercept-PC only), untreated HPL, and FBS. MSC morphology, doubling time, immunophenotype, immunosuppressive activity, and differentiation capacity were compared. RESULTS:Expanded cells had typical spindle morphology and showed higher viability in all HPL conditions than in FBS. The DVI-HPL and FBS-expanded cells were morphologically larger than in I-HPL and HPL supplements. The cumulative population doubling was lower using DVI-HPL than with HPL and I-HPL, but significantly higher than using FBS. Immunophenotype was not affected by the supplements used. Immunosuppressive activity was maintained with all supplements. Differentiation capacity into chondrocytes and osteocytes was more effective in DVI-HPL but less toward adipocytes compared to other supplements. CONCLUSIONS: Human PLT lysate made fromIntercept-PCs subjected to S/D treatment may be an alternative to untreated HPL and to I-HPL for BM-MSC expansion. This finding reinforces the potential of HPL as a virally safe alternative to FBS for clinical grade MSC expansion protocols. ABBREVIATIONS: BDNF = brain-derived neurotrophic factor; BM-MSC(s) = bone marrow-derived mesenchymal stromal cells; BrdU = bromodeoxyuridine/5-bromo-2 0 -deoxyuridine; CCK-8 = cell counting kit-8; DVI = double-virally-inactivated; DVI-HPL = doublevirally-inactivated (intercept-solvent/detergent) human platelet lysate; ECM = extracellular matrix; EGF = epidermal growth factor FBS = fetal bovine serum; HGF = hepatocyte growth factor HLA-DR = human leukocyte antigen DR; HPL = human platelet lysate; IGF = insulin-like growth factor-1; I-HPL = Intercept human platelet lysate; MSC(s) = mesenchymal stromal cells; PAS = platelet additive solution; PB19V = parvovirus B19; PC(s) = platelet concentrate(s); PD (s) = population doubling(s); PDGF-AB = platelet-derived growth factor-AB; PF4 = platelet factor 4; PHA = phytohemagglutinin; PPARG = peroxisome proliferator-activated receptor gamma; SOX9 = SRY (sex-determining region Y)-box 9; SPP1 = secreted phosphoprotein 1. From the

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Section: Discussionsupporting

confidence: 92%

A double‐virally‐inactivated (Intercept–solvent/detergent) human platelet lysate for in vitro expansion of human mesenchymal stromal cells

Barro

Nebie

et al. 2019

Transfusion

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“…In terms of content, one major difference between the two human nutritive substitutes used in the present study is the presence in hPL of plasma fibrinogen and of heparin, which is required to avoid gelation of the medium. In light of previous studies suggesting that the heparin concentration may affect stem cell proliferation and/or differentiation [ 70 ], we investigated the effects of heparin on myogenic commitment in MuStem cells. hMuStem cells HS were cultured in the presence of increasing doses of heparin (0.5–5 IU/ml), with the highest dose corresponding to that found in hPL-GM.…”

Section: Resultsmentioning

confidence: 99%

Human serum and platelet lysate are appropriate xeno-free alternatives for clinical-grade production of human MuStem cell batches

et al. 2018

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BackgroundCanine MuStem cells have demonstrated regenerative efficacy in a dog model of muscular dystrophy, and the recent characterization of human counterparts (hMuStem) has highlighted the therapeutic potential of this muscle-derived stem cell population. To date, these cells have only been generated in research-grade conditions. However, evaluation of the clinical efficacy of any such therapy will require the production of hMuStem cells in compliance with good manufacturing practices (GMPs). Because the current use of fetal bovine serum (FBS) to isolate and expand hMuStem cells raises several ethical, safety, and supply concerns, we assessed the use of two alternative xeno-free blood derivatives: human serum (HS) and a human platelet lysate (hPL).MethodshMuStem cells were isolated and expanded in vitro in either HS-supplemented or hPL-supplemented media and the proliferation rate, clonogenicity, myogenic commitment potential, and oligopotency compared with that observed in FBS-supplemented medium. Flow cytometry and high-throughput 3′-digital gene expression RNA sequencing were used to characterize the phenotype and global gene expression pattern of hMuStem cells cultured with HS or hPL.ResultsHS-supplemented and hPL-supplemented media both supported the isolation and long-term proliferation of hMuStem cells. Compared with FBS-based medium, both supplements enhanced clonogenicity and allowed for a reduction in growth factor supplementation. Neither supplement altered the cell lineage pattern of hMuStem cells. In vitro differentiation assays revealed a decrease in myogenic commitment and in the fusion ability of hMuStem cells when cultured with hPL. In return, this reduction of myogenic potential in hPL-supplemented cultures was rapidly reversed by substitution of hPL with HS or fibrinogen-depleted hPL. Moreover, culture of hMuStem cells in hPL hydrogel and fibrinogen-depleted hPL demonstrated that myogenic differentiation potential is maintained in heparin-free hPL derivatives.ConclusionsOur findings indicate that HS and hPL are efficient and viable alternatives to FBS for the preparation of hMuStem cell batches in compliance with GMPs.Electronic supplementary materialThe online version of this article (10.1186/s13287-018-0852-y) contains supplementary material, which is available to authorized users.

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“…The effects of pathogen inactivation on growth factor stability during storage have previously been studied. It was demonstrated that UVC treatment of platelet concentrates had no effect on concentrations of EGF, FGF-2, PDGF-AB, VEGF, or insulin-like growth factor (IGF) [35]. In a study specifically conducted on the INTERCEPT™ Blood System…”

Section: Discussionmentioning

confidence: 99%

Effects of amotosalen treatment on human platelet lysate bioactivity: A proof-of-concept study

2020

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Background Clinical application of mesenchymal stromal cells (MSCs) usually requires an in vitro expansion step to reach clinically relevant numbers. In vitro cell expansion necessitates supplementation of basal mammalian cell culture medium with growth factors. To avoid using supplements containing animal substances, human platelet lysates (hPL) produced from expired and pathogen inactivated platelet concentrates can be used in place of fetal bovine serum. However, globally, most transfusion units are currently not pathogen inactivated. As blood banks are the sole source of platelet concentrates for hPL production, it is important to ensure product safety and standardized production methods. In this proof-of-concept study we assessed the feasibility of producing hPL from expired platelet concentrates with pathogen inactivation applied after platelet lysis by evaluating the retention of growth factors, cytokines, and the ability to support MSC proliferation and tri-lineage differentiation. Methodology/Principal findings Bone marrow-derived MSCs (BM-MSCs) were expanded and differentiated using hPL derived from pathogen inactivated platelet lysates (hPL-PIPL), with pathogen inactivation by amotosalen/ultraviolet A treatment applied after lysis of expired platelets. Results were compared to those using hPL produced from conventional expired pathogen inactivated platelet concentrates (hPL-PIPC), with pathogen inactivation applied after blood donation. hPL-PIPL treatment had lower concentrations of soluble growth factors and cytokines than hPL-PIPC treatment. When used as supplementation in cell culture, BM-MSCs proliferated at a reduced rate, but more consistently, in hPL-PIPL than in hPL-PIPC. The ability to support tri-lineage differentiation was comparable between lysates. Conclusion/Significance These results suggest that functional hPL can be produced from expired and untreated platelet lysates by applying pathogen inactivation after platelet lysis. When carried out post-PLOS ONE

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Pathogen reduction through additive-free short-wave UV light irradiation retains the optimal efficacy of human platelet lysate for the expansion of human bone marrow mesenchymal stem cells

Cited by 30 publications

References 72 publications

A double‐virally‐inactivated (Intercept–solvent/detergent) human platelet lysate for in vitro expansion of human mesenchymal stromal cells

A double‐virally‐inactivated (Intercept–solvent/detergent) human platelet lysate for in vitro expansion of human mesenchymal stromal cells

Human serum and platelet lysate are appropriate xeno-free alternatives for clinical-grade production of human MuStem cell batches

Effects of amotosalen treatment on human platelet lysate bioactivity: A proof-of-concept study

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