Human platelet lysate is a feasible candidate to replace fetal calf serum as medium supplement for blood vascular and lymphatic endothelial cells

Purpose: Corneal endothelial cell (CEC) therapy can be used as a promising therapeutic option for patients with various corneal endothelial dysfunctions. In this study, we compared the proliferative effect of human platelet lysate (HPL), as a xeno-free medium supplement, with Y-27632 Rho/rho-associated protein kinase (ROCK) inhibitor, as a wellknown proliferative and adhesive agent for CECs, and fetal bovine serum (FBS) as the control, in the culture medium of human corneal endothelial cells (HCECs). Methods: We isolated HCECs from human donors and treated the cells as three different treatment groups including 20% HPL only, 10 μM Y-27632 ROCK inhibitor, combination of 20% HPL and 10 μM Y-27632 ROCK inhibitor, and 20% FBS as the control group. ELISA cell proliferation assay and cell counting was performed on the treated cells. Finally, HCECs were characterized by morphology and immunocytochemistry (ICC). Results: There was no significant proliferative effect of HPL on cell proliferation compared with the cells treated with Y-27632 ROCK inhibitor or the combination of HPL and Y-27632 ROCK inhibitor, but all the respected treatments had significant inducible effect on cell proliferation as compared with FBS-treated cells. The cells grown in all three treatment groups exhibited CEC morphology. Also, there was a higher expression of Na+/K+-ATPase and ZO-1, as CEC characteristic markers, in the culture of HCECs treated with HPL as compared with FBS. Conclusion: HPL offers a xeno−free and affordable medium supplement for CEC expansion that can be used in clinical applications.

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Abstract 3100: Replacing fetal calf serum by human platelet lysate in cancer research and toxicology

10.1158/1538-7445.am2018-3100 ◽

2018 ◽

Author(s):

Oliver H. Krämer

Keyword(s):

Cancer Research ◽

Fetal Calf Serum ◽

Human Platelet ◽

Calf Serum ◽

Platelet Lysate ◽

Human Platelet Lysate

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Proliferation and Differentiation of Intestinal Caco-2 Cells Are Maintained in Culture with Human Platelet Lysate Instead of Fetal Calf Serum

Cells ◽

10.3390/cells10113038 ◽

2021 ◽

Vol 10 (11) ◽

pp. 3038

Author(s):

Dalanda Wanes ◽

Hassan Y. Naim ◽

Franziska Dengler

Keyword(s):

Cell Viability ◽

Fetal Calf Serum ◽

Human Platelet ◽

Calf Serum ◽

Platelet Lysate ◽

Model Systems ◽

Structural Protein ◽

Cell Extracts ◽

Gene And Protein Expression ◽

Human Platelet Lysate

Cell lines are widely used as in vitro model systems and substitute for animal experiments. The frequently used Caco-2 cell line is considered to reflect characteristics of differentiated intestinal epithelium. However, the need to culture the cells with fetal calf serum (FCS) induces a high variability, risk of contamination and is ethically disputed. We tested the culture of Caco-2 cells with human platelet lysate (PL) instead of FCS. We compared cell viability and differentiation by measuring ATP levels, gene and protein expression of specific markers in total cell extracts, brush border membrane vesicles (BBM) and lipid rafts (LR). Cell viability was slightly enhanced in cells grown with PL compared to FCS. The cells differentiated to an intestinal phenotype like the cells cultured in FCS, as indicated by the similar gene expression levels of hexose and protein transport proteins and the structural protein VILLIN. BBM showed a comparable distribution of the intestinal hydrolases, indicating a maintained cell membrane polarity. The distribution of the marker protein FLOTILLIN-2 in LR was also similar. We conclude that PL is an exquisite and suitable replacement for FCS in the culture of Caco-2 cells that can eliminate many disadvantages incurred due to the use of FCS.

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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

Cells ◽

10.3390/cells9040918 ◽

2020 ◽

Vol 9 (4) ◽

pp. 918 ◽

Cited By ~ 3

Author(s):

Maria Karadjian ◽

Anne-Sophie Senger ◽

Christopher Essers ◽

Sebastian Wilkesmann ◽

Raban Heller ◽

...

Keyword(s):

Bone Marrow ◽

Osteogenic Differentiation ◽

Fetal Calf Serum ◽

Culture Media ◽

Human Platelet ◽

Calf Serum ◽

Calcium Content ◽

Platelet Lysate ◽

Protein Source ◽

Human Platelet Lysate

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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In vitro expansion of human primary endothelial cells for clinical use using EndoGo™ XF Medium supplemented with PLTGold ® human platelet lysate

Cytotherapy ◽

10.1016/j.jcyt.2018.02.246 ◽

2018 ◽

Vol 20 (5) ◽

pp. S87

Author(s):

V. Alonso-Camino ◽

B. Mirsch

Keyword(s):

Endothelial Cells ◽

Human Platelet ◽

Platelet Lysate ◽

Clinical Use ◽

In Vitro Expansion ◽

Human Platelet Lysate

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Human platelet lysate as a potential clinical-translatable supplement to support the neurotrophic properties of human adipose-derived stem cells

Stem Cell Research & Therapy ◽

10.1186/s13287-020-01949-4 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Silvia Palombella ◽

Martino Guiotto ◽

Gillian C. Higgins ◽

Laurent L. Applegate ◽

Wassim Raffoul ◽

...

Keyword(s):

Stem Cells ◽

Growth Factor ◽

Peripheral Nerve ◽

Culture Medium ◽

Human Platelet ◽

Platelet Lysate ◽

Clinical Translation ◽

Adipose Derived Stem Cells ◽

Medium Supplement ◽

Human Platelet Lysate

Abstract Background The autologous nerve graft, despite its donor site morbidity and unpredictable functional recovery, continues to be the gold standard in peripheral nerve repair. Rodent research studies have shown promising results with cell transplantation of human adipose-derived stem cells (hADSC) in a bioengineered conduit, as an alternative strategy for nerve regeneration. To achieve meaningful clinical translation, cell therapy must comply with biosafety. Cell extraction and expansion methods that use animal-derived products, including enzymatic adipose tissue dissociation and the use of fetal bovine serum (FBS) as a culture medium supplement, have the potential for transmission of zoonotic infectious and immunogenicity. Human-platelet-lysate (hPL) serum has been used in recent years in human cell expansion, showing reliability in clinical applications. Methods We investigated whether hADSC can be routinely isolated and cultured in a completely xenogeneic-free way (using hPL culture medium supplement and avoiding collagenase digestion) without altering their physiology and stem properties. Outcomes in terms of stem marker expression (CD105, CD90, CD73) and the osteocyte/adipocyte differentiation capacity were compared with classical collagenase digestion and FBS-supplemented hADSC expansion. Results We found no significant differences between the two examined extraction and culture protocols in terms of cluster differentiation (CD) marker expression and stem cell plasticity, while hADSC in hPL showed a significantly higher proliferation rate when compared with the usual FBS-added medium. Considering the important key growth factors (particularly brain-derived growth factor (BDNF)) present in hPL, we investigated a possible neurogenic commitment of hADSC when cultured with hPL. Interestingly, hADSC cultured in hPL showed a statistically higher secretion of neurotrophic factors BDNF, glial cell-derived growth factor (GDNF), and nerve-derived growth factor (NFG) than FBS-cultured cells. When cocultured in the presence of primary neurons, hADSC which had been grown under hPL supplementation, showed significantly enhanced neurotrophic properties. Conclusions The hPL-supplement medium could improve cell proliferation and neurotropism while maintaining stable cell properties, showing effectiveness in clinical translation and significant potential in peripheral nerve research.

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