A highly standardized and characterized human platelet lysate for efficient and reproducible expansion of human bone marrow mesenchymal stromal cells

Viau, Sabrina; Lagrange, Anaïs; Chabrand, Lucie; Lorant, Judith; Charrier, Marine; Rouger, Karl; Alvarez, I.; Eap, Sandy; Delorme, Bruno

doi:10.1016/j.jcyt.2019.04.053

Cited by 31 publications

(41 citation statements)

References 69 publications

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“…hPL is generally produced out of a varying number of pooled human platelet concentrates [13,18,[42][43][44][45] after platelet lysis by freezing-thawing cycles [20,[43][44][45] or activation by thrombin [13,42] to liberate the substances required for cell culture that are stored in platelet granules [14,15]. However, there is no good manufacturing practice statement concerning the platelet lysis method in hPL production [13].…”

Section: Discussionmentioning

confidence: 99%

“…The most common alternative to FCS is human platelet lysate (hPL) [1], which is produced from human platelets and contains proteins and growth factors necessary for, inter alia, in-vitro human bone-marrow-derived mesenchymal stromal cell (BMSC) expansion [13][14][15][16][17][18] and osteogenic differentiation [15,16,19]. hPL products can have smaller batch-to-batch variabilities by pooling multiple batches of platelet concentrates, hence extensive pre-testing becomes obsolete [17,20].…”

Section: Introductionmentioning

confidence: 99%

“…hPL products can have smaller batch-to-batch variabilities by pooling multiple batches of platelet concentrates, hence extensive pre-testing becomes obsolete [17,20]. It has already been shown that hPL-supplemented expansion medium (ESM) augments the proliferation rates of BMSC [11,[16][17][18][21][22][23][24][25][26][27][28][29], and does not affect BMSC properties as part of ESM [21]. Several studies evaluated BMSC differentiation potential after cell expansion in hPL-containing medium, while the differentiation medium itself contained only FCS as a supplement [17,18,22,23,[28][29][30].…”

Section: Introductionmentioning

confidence: 99%

“…It has already been shown that hPL-supplemented expansion medium (ESM) augments the proliferation rates of BMSC [11,[16][17][18][21][22][23][24][25][26][27][28][29], and does not affect BMSC properties as part of ESM [21]. Several studies evaluated BMSC differentiation potential after cell expansion in hPL-containing medium, while the differentiation medium itself contained only FCS as a supplement [17,18,22,23,[28][29][30]. There have also been multiple studies reporting that hPL does not affect osteogenic differentiation [16,17,21,22,24,[27][28][29]31] or even increases the osteogenic differentiation potential of BMSC [22,23,25,26].…”

Section: Introductionmentioning

confidence: 99%

“…However, it remains unclear how hPL as a part of an osteogenic differentiation medium (ODM) influences the kinetics of the osteogenic differentiation of BMSC. Furthermore, the necessary concentrations of hPL reported differ from 2% to 10% [18,20,25,26,[29][30][31][32][33][34], pointing out that there is no established standard protocol for hPL supplementation in ODM for BMSC yet, although the concentration of serum supplement in media resulting in different concentrations of growth factors and other components of the serum influences cells in culture [1].…”

Section: Introductionmentioning

confidence: 99%

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Human Platelet Lysate Can Replace Fetal Calf Serum as a Protein Source to Promote Expansion and Osteogenic Differentiation of Human Bone-Marrow-Derived Mesenchymal Stromal Cells

Karadjian

Senger

Essers

et al. 2020

Cells

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Fetal calf serum (FCS) is frequently used as a growth factor and protein source in bone-marrow-derived mesenchymal stromal cell (BMSC) culture media, although it is a xenogenic product presenting multiple disadvantages including but not limited to ethical concerns. A promising alternative for FCS is human platelet lysate (hPL), which is produced out of human platelet concentrates and happens to be a stable and reliable protein source. In this study, we investigated the influence of hPL in an expansion medium (ESM) and an osteogenic differentiation medium (ODM) on the proliferation and osteogenic differentiation capacity of human BMSC. Therefore, we assessed population doublings during cell expansion, performed alizarin red staining to evaluate the calcium content in the extracellular matrix and determined the activity of alkaline phosphatase (ALP) as osteogenic differentiation correlates. The proliferation rate of BMSC cultured in ESM supplemented with hPL exceeded the proliferation rate of BMSC cultured in the presence of FCS. Furthermore, the calcium content and ALP activity was significantly higher in samples incubated in hPL-supplemented ODM, especially in the early phases of differentiation. Our results show that hPL can replace FCS as a protein supplier in cell culture media and does not negatively affect the osteogenic differentiation capacity of BMSC.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Human Platelet Lysate Can Replace Fetal Calf Serum as a Protein Source to Promote Expansion and Osteogenic Differentiation of Human Bone-Marrow-Derived Mesenchymal Stromal Cells

Karadjian

Senger

Essers

et al. 2020

Cells

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Multifunctional Surfaces for Improving Soft Tissue Integration

Vilaça

Domingues

Tiainen

et al. 2021

Adv Healthcare Materials

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Metallic implants are widely used in diverse clinical applications to aid in recovery from lesions or to replace native hard tissues. However, the lack of integration of metallic surfaces with soft tissue interfaces causes the occurrence of biomaterial-associated infections, which can trigger a complicated inflammatory response and, ultimately, implant failure. Here, a multifunctional implant surface showing nanoscale anisotropy, based on the controlled deposition of cellulose nanocrystals (CNC), and biological activity derived from platelet lysate (PL) biomolecules sequestered and presented on CNC surface, is proposed. The anisotropic radial nanopatterns are produced on polished titanium surfaces by spin-coating CNC at high speed. Furthermore, CNC surface chemistry allows to further sequester and form a coating of bioactive molecules derived from PL. The surface anisotropy provided by CNC guides fibroblasts growth and alignment up to 14 days of culture. Moreover, PL-derived biomolecules polarize macrophages toward the M2-like anti-inflammatory phenotype. These results suggest that the developed multifunctional surfaces can promote soft tissue integration to metallic implants and, at the same time, prevent bacterial invasion, tissue inflammation, and failure of biomedical metallic implants.

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The proteomic and particle composition of human platelet lysate for cell therapy products

Rodrigues

Valim

Berger

et al. 2022

J of Cellular Biochemistry

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Following health agencies warning, the use of animal origin supplements should be avoided in biological products proposed as therapy in humans.Platelet lysate and several other growth factors sources are alternatives to replace fetal calf serum, the current gold standard in clinical-grade cell culture. However, the platelet supplement's content lacks data due to different production methods. The principle behind these products relays on the lysis of platelets that release several proteins, some of which are contained in heterogeneous granules and coordinate biological functions. This study aims to analyze the composition and reproducibility of a platelet lysate produced with a standardized method, by describing several batches' protein and particle content using proteomics and dynamic light scattering. Proteomics data revealed a diversified protein content, with some related to essential cellular processes such as proliferation, morphogenesis, differentiation, biosynthesis, adhesion, and metabolism. It also detected proteins responsible for activation and binding of transforming growth factor beta, hepatocyte growth factor, and insulin-like growth factor. Total protein, biochemical, and growth factors quantitative data showed consistent and reproducible values across batches. Novel data on two major particle populations is presented, with high dispersion level at 231 ± 96 d.nm and at 30 ± 8 d.nm, possibly being an important way of protein trafficking through the cellular microenvironment. This experimental and descriptive analysis aims to support the content definition and quality criteria of a cell supplement for clinical applications.

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A highly standardized and characterized human platelet lysate for efficient and reproducible expansion of human bone marrow mesenchymal stromal cells

Cited by 31 publications

References 69 publications

Human Platelet Lysate Can Replace Fetal Calf Serum as a Protein Source to Promote Expansion and Osteogenic Differentiation of Human Bone-Marrow-Derived Mesenchymal Stromal Cells

Human Platelet Lysate Can Replace Fetal Calf Serum as a Protein Source to Promote Expansion and Osteogenic Differentiation of Human Bone-Marrow-Derived Mesenchymal Stromal Cells

Multifunctional Surfaces for Improving Soft Tissue Integration

The proteomic and particle composition of human platelet lysate for cell therapy products

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