Donor Age of Human Platelet Lysate Affects Proliferation and Differentiation of Mesenchymal Stem Cells

Abstract

The regenerative potential declines upon aging. This might be due to cell-intrinsic changes in stem and progenitor cells or to influences by the microenvironment. Mesenchymal stem cells (MSC) raise high hopes in regenerative medicine. They are usually culture expanded in media with fetal calf serum (FCS) or other serum supplements such as human platelet lysate (HPL). In this study, we have analyzed the impact of HPL-donor age on culture expansion. 31 single donor derived HPLs (25 to 57 years old) were simultaneously compared for culture of MSC. Proliferation of MSC did not reveal a clear association with platelet counts of HPL donors or growth factors concentrations (PDGF-AB, TGF-β1, bFGF, or IGF-1), but it was significantly higher with HPLs from younger donors (<35 years) as compared to older donors (>45 years). Furthermore, HPLs from older donors increased activity of senescence-associated beta-galactosidase (SA-βgal). HPL-donor age did not affect the fibroblastoid colony-forming unit (CFU-f) frequency, immunophenotype or induction of adipogenic differentiation, whereas osteogenic differentiation was significantly lower with HPLs from older donors. Concentrations of various growth factors (PDGF-AB, TGF-β1, bFGF, IGF-1) or hormones (estradiol, parathormone, leptin, 1,25 vitamin D3) were not associated with HPL-donor age or MSC growth. Taken together, our data support the notion that aging is associated with systemic feedback mechanisms acting on stem and progenitor cells, and this is also relevant for serum supplements in cell culture: HPLs derived from younger donors facilitate enhanced expansion and more pronounced osteogenic differentiation.

Venous Blood Derivatives as FBS-Substitutes for Mesenchymal Stem Cells: A Systematic Scoping Review

“…VBD concentrations added to culture medium ranged from 0.5 to 30% (54)(55)(56). Overall, VBD allowed for either, maintenance of MCS's fibroblast-like morphology (17,43,57), high proliferation (58-60), high colony-formation ability (52,61,62) and maintenance of multipotency (63)(64)(65)(66)(67). A linear dose-dependent response regarding medium concentration was not observed for evaluated VBD (54,56,68).…”

“…In the protocol to obtain the HPL, the Induction of fibrin clotting formation with calcium chloride (instead of thrombin) and posterior centrifugation provides a serum with the same profile of growing factor and cytokines than conventional HPL activated with thrombin, resulting in a serum without free of xenogeneic substances such as porcine thrombin (118). HPL supplemented without anticoagulants tend to form a translucent and viscous gel in 1 h providing a natural scaffold for MSC culture and expansion (61). This matrix, composed by a fibrin network, was biocompatible and biodegradable.…”

“…This matrix, composed by a fibrin network, was biocompatible and biodegradable. Additionally, MSC cultured in this fibrin matrix presented higher proliferation since growth is available in three-dimensional environment, increasing the culture area (8,61).…”

Partnership between platelet-rich plasma and mesenchymal stem cells: in vitro experience

“…When those steps are performed, they proceed to filtration through 0.22 um filters and addition of heparin (2U/ml) to the lysates 35,40,57 . This procedure is the most commonly found in the papers studied, but there are some authors which prefer performing chemical activation instead of thermic/mechanical activation and use of thrombin and/or calcium chloride 7,17,34,41,45,49 .…”

Platelet-functionalized three-dimensional poly-&epsilon;-caprolactone fibrous scaffold prepared using centrifugal spinning for delivery of growth factors

“…The allogenic preparations based on buffy coat also enable elimination of donor-to-donor variations by preparing mixed platelet isolates from multiple donors. The importance of the problem was highlighted by Lohmann et al, 38 who observed relationship between patient age and proliferation of MSCs.…”