Systematic review and meta-analysis on the use of human platelet lysate for mesenchymal stem cell cultures: comparison with fetal bovine serum and considerations on the production protocol

Palombella, Silvia; Orfei, Carlotta Perucca; Castellini, Greta; Gianola, Silvia; Lopa, Silvia; Mastrogiacomo, Maddalena; Moretti, Matteo; Girolamo, Laura de

doi:10.1186/s13287-022-02815-1

Cited by 19 publications

(13 citation statements)

References 64 publications

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“…In addition, others demonstrated that platelets or platelet-containing preparations can be used to promote the regenerative potential of human AD-MSC ( 30 ) and several studies have reported an improved therapeutic efficacy of MSC from different sources induced by platelet rich plasma ( 31 – 33 ). After numerous promising studies have reported the preservation of the basic characteristics of human BM- and AD-MSC with hPL ( 34 – 37 ), and functional studies using hPL ( 38 , 39 ) have also been carried out in human medicine, the time has come to perform functional assays for ePL as well.…”

Section: Introductionmentioning

confidence: 99%

Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate

Hagen

Niebert²,

Brandt³

et al. 2022

Front. Vet. Sci.

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Successful translation of multipotent mesenchymal stromal cell (MSC)-based therapies into clinical reality relies on adequate cell production procedures. These should be available not only for human MSC, but also for MSC from animal species relevant to preclinical research and veterinary medicine. The cell culture medium supplementation is one of the critical aspects in MSC production. Therefore, we previously established a scalable protocol for the production of buffy-coat based equine platelet lysate (ePL). This ePL proved to be a suitable alternative to fetal bovine serum (FBS) for equine adipose-derived (AD-) MSC culture so far, as it supported AD-MSC proliferation and basic characteristics. The aim of the current study was to further analyze the functional properties of equine AD-MSC cultured with the same ePL, focusing on cell fitness, genetic stability and pro-angiogenic potency. All experiments were performed with AD-MSC from n = 5 horses, which were cultured either in medium supplemented with 10% FBS, 10% ePL or 2.5% ePL. AD-MSC cultured with 2.5% ePL, which previously showed decreased proliferation potential, displayed higher apoptosis but lower senescence levels as compared to 10% ePL medium (p < 0.05). Non-clonal chromosomal aberrations occurred in 8% of equine AD-MSC cultivated with FBS and only in 4.8% of equine AD-MSC cultivated with 10% ePL. Clonal aberrations in the AD-MSC were neither observed in FBS nor in 10% ePL medium. Analysis of AD-MSC and endothelial cells in an indirect co-culture revealed that the ePL supported the pro-angiogenic effects of AD-MSC. In the 10% ePL group, more vascular endothelial growth factor (VEGF-A) was released and highest VEGF-A concentrations were reached in the presence of ePL and co-cultured cells (p < 0.05). Correspondingly, AD-MSC expressed the VEGF receptor-2 at higher levels in the presence of ePL (p < 0.05). Finally, AD-MSC and 10% ePL together promoted the growth of endothelial cells and induced the formation of vessel-like structures in two of the samples. These data further substantiate that buffy-coat-based ePL is a valuable supplement for equine AD-MSC culture media. The ePL does not only support stable equine AD-MSC characteristics as demonstrated before, but it also enhances their functional properties.

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Section: Introductionmentioning

confidence: 99%

Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate

Hagen

Niebert²,

Brandt³

et al. 2022

Front. Vet. Sci.

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“…This raises quite serious concerns given the increasing use of PL as a substitute for FBS (15,39). Recent systematic reviews of studies comparing PL and FBS for MSC expansion have reported no signi cant differences in immunosuppression (22,23), however, the differences observed in this study, particularly between PL and PS2 are stark. One potential reason for enhanced immunosuppression in MSCs cultured in PS2 could be that this effect is extracellular vesicle (EV)-mediated.…”

Section: Discussionmentioning

confidence: 51%

“…These have predominantly compared one selected media to the gold standard of 10% FBS. PL has been the most commonly studied supplement and has been the subject of many comparisons with FBS medium, the results of which have been analysed in recent systematic reviews (21)(22)(23). Comprehensive studies comparing PS2 medium to standard FBS-containing medium both with and without additional supplementation with FGF2 for the isolation and expansion of MSCs have previously been performed (20,24).…”

Section: Introductionmentioning

confidence: 99%

Media Matters: Culture Medium-Dependent Hypervariable Phenotype of Mesenchymal Stromal Cells

Fitzgerald,

Shaw,

Murphy

et al. 2023

Preprint

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Background Despite a long history of investigation and sustained efforts in clinical testing, the number of market authorisations for mesenchymal stromal cell (MSC) therapies remains limited, with none approved by the United States Food and Drug Administration (FDA). Several barriers are impeding the clinical progression of MSC therapies, to the forefront of these is a lack of standardised manufacturing protocols which is further compounded by an absence of biologically meaningful characterisation and release assays. A look at clinical trial registries demonstrates the diversity of MSC expansion protocols with variabilities in cell source, isolation method and expansion medium, amongst other culture variables, making it extraordinarily difficult to compare study outcomes. Current identification and characterisation standards are insufficient; they are not specific to MSCs and do not indicate cell function or therapeutic action. Methods This work analysed the influence of five widely used culture media formulations on the colony forming potential, proliferation kinetics, trilineage differentiation potential and immunomodulatory potential of human bone marrow derived MSCs (BM-MSCs). The surface marker expression profiles were also characterised using a high-content flow cytometry screening panel of 243 markers. Results Significant differences in the biological attributes of BM-MSCs including clonogenicity, proliferation, differentiation propensity and immunomodulatory capacity were revealed in response to the composition of the culture medium. Despite their biological differences, all cell preparations uniformly and strongly expressed the standard positive markers proposed for BM-MSCs; CD73, CD90 and CD105. Immunophenotypic profiling revealed that the culture medium also had a significant influence on the surface proteome, with one third of tested markers exhibiting variable expression profiles. Principal component analysis demonstrated that BM-MSCs isolated and expanded in a proprietary xeno- and serum-free medium displayed the most consistent cell phenotypes with little variability between donors compared to platelet lysate (PL) and fetal bovine serum (FBS)-containing media. Conclusions These data suggest that media composition has a highly significant impact on the biological attributes of MSCs, but standard surface marker tests conceal these differences. The results indicate a need for (1) standardised approaches to manufacturing, with an essential focus on defined media and (2) new biologically relevant tests for MSC characterisation and product release.

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“…PL was obtained by freeze-thawing PRP samples obtained from the AU Ibni Sina Hospital Blood Center (AU ISHBC # 13-625-16). PRP samples from five healthy donors were pooled to reduce inter-donor variability [23]. Pooled PRP was kept at −80 • C for 24 h and then incubated for 1 h in a 37 • C water bath.…”

Section: Preparation Of Plmentioning

confidence: 99%

Biomimetic mineralization of platelet lysate/oxidized dextran cryogel as a macroporous 3D composite scaffold for bone repair

Şeker,

Aral,

Elçin

et al. 2024

Biomed. Mater.

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Scaffold development approaches using autologous sources for tissue repair are of great importance in obtaining bio-active/-compatible constructs. Platelet-rich plasma (PRP) containing various growth factors and platelet lysate (PL) derived from PRP are autologous products that have the potential to accelerate the tissue repair response by inducing a transient inflammatory event. Considering the regenerative capacity of PRP and PL, PRP/PL-based scaffolds are thought to hold great promise in tissue engineering by providing an ideal autologous growth factor source and mechanical support for cells. Here, a bio-mineralized PRP-based scaffold was developed using oxidized dextran (OD) and evaluated for future application in bone tissue engineering. Prepared PL/OD scaffolds were incubated in simulated body fluid (SBF) for 7, 14 and 21 day periods. Mineralized PL/OD scaffolds were characterized using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction spectroscopy (XRD), scanning electron microscopy (SEM), thermogravimetric analysis (TGA), porosity and compression tests. SEM and EDX analyses revealed mineral accumulation on the PL/OD scaffold as a result of SBF incubation. In vitro cytotoxicity and in vitro hemolysis tests revealed that the scaffolds were non-toxic and hemocompatible. Additionally, human osteoblasts (hOBs) exhibited good attachment and spreading behavior on the scaffolds and maintained their viability throughout the culture period. The alkaline phosphatase activity assay and calcium release results revealed that PL/OD scaffolds preserved the osteogenic properties of hOBs. Overall, findings suggest that mineralized PL/OD scaffold may be a promising scaffold for bone tissue engineering.

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Systematic review and meta-analysis on the use of human platelet lysate for mesenchymal stem cell cultures: comparison with fetal bovine serum and considerations on the production protocol

Cited by 19 publications

References 64 publications

Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate

Functional properties of equine adipose-derived mesenchymal stromal cells cultured with equine platelet lysate

Media Matters: Culture Medium-Dependent Hypervariable Phenotype of Mesenchymal Stromal Cells

Biomimetic mineralization of platelet lysate/oxidized dextran cryogel as a macroporous 3D composite scaffold for bone repair

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