Human platelet lysate in mesenchymal stromal cell expansion according to a GMP grade protocol: a cell factory experience

Abstract

BackgroundThe use of platelet lysate (PL) for the ex-vivo expansion of mesenchymal stromal/stem cells (MSCs) was initially proposed by Doucet et al. in 2005, as an alternative to animal serum. Moreover, regulatory authorities discourage the use of fetal bovine serum (FBS) or other animal derivatives, to avoid risk of zoonoses and xenogeneic immune reactions. Even if many studies investigated PL composition, there still are some open issues related to its use in ex-vivo MSC expansion, especially according to good manufacturing practice (GMP) grade protocols.MethodsAs an authorized cell factory, we report our experience using standardized PL produced by Azienda Ospedaliero Universitaria Meyer Transfusion Service for MSC expansion according to a GMP grade clinical protocol. As suggested by other authors, we performed an in-vitro test on MSCs versus MSCs cultured with FBS that still represents the best way to test PL batches. We compared 12 MSC batches cultured with DMEM 5% PL with similar batches cultured with DMEM 10% FBS, focusing on the MSC proliferation rate, MSC surface marker expression, MSC immunomodulatory and differentiation potential, and finally MSC relative telomere length.ResultsResults confirmed the literature data as PL increases cell proliferation without affecting the MSC immunophenotype, immunomodulatory potential, differentiation potential and relative telomere length.ConclusionsPL can be considered a safe alternative to FBS for ex-vivo expansion of MSC according to a GMP grade protocol. Our experience confirms the literature data: a large number of MSCs for clinical applications can be obtained by expansion with PL, without affecting the MSC main features. Our experience underlines the benefits of a close collaboration between the PL producers (transfusion service) and the end users (cell factory) in a synergy of skills and experiences that can lead to standardized PL production.Electronic supplementary materialThe online version of this article (10.1186/s13287-018-0863-8) contains supplementary material, which is available to authorized users.

Autologous adipose-derived stem cells for the treatment of complex cryptoglandular perianal fistula: A randomized clinical trial with long-term follow-up

“…Unlike in other studies, here ASCs were expanded using human platelet lysate in the culture media, which may have the advantage of avoiding the use of animal‐derived factors. Human platelet lysate is widely used to grow MSCs, and many studies have demonstrated that this confers a degree of safety that is at least equivalent to that offered by fetal bovine serum . Indeed, some studies have suggested that the use of platelet lysate may increase cell yield and shorten the time for ex vivo expansion without epigenetic alterations …”

“…Human platelet lysate is widely used to grow MSCs, [23][24][25] and many studies have demonstrated that this confers a degree of safety that is at least equivalent to that offered by fetal bovine serum. 26,27 Indeed, some studies have suggested that the use of platelet lysate may increase cell yield and shorten the time for ex vivo expansion without epigenetic alterations. 25,27 One of the issues regarding the use of somatic cell medicinal products has been the selection of the cell dose.…”

Human platelet lysate-based nanocomposite bioink for bioprinting hierarchical fibrillar structures

“…[14,29] This is reflected in the significantly higher metabolic activity of cells in HUink compared to the other two bioink materials (figure 5(c)). In addition, although in this study pooled PL batches were prepared in house, the increasing trend on the use of standardized clinical grade human PL as xeno-free alternative to animal-derived serum in cell culture, [30,31] represents an advantage over similar materials such as ECM bioinks (e.g. GelMA or decellurarised matrix), [11,32] which are typically of xenogenic origin, a significant barrier for clinical translation, and present high variability on the biochemical, topological and viscoelastic properties depending on the decellularization protocol.…”

Modeling Biological and Economic Uncertainty on Cell Therapy Manufacturing: the Choice of Culture Media Supplementation

“…Quality controls for release testing of MSCs are related to safety, identity, purity and potency [61,62]. While some studies claim that hPL does not modify the immunomodulatory characteristics and the multipotency of MSCs [28,58,60,63], others show that there is a decrease in the immunomodulatory activity and surface marker modifications in BM-MSCs cultured with hPL [9,40,64].…”