scholarly journals Effects of amotosalen treatment on human platelet lysate bioactivity

2019 ◽  
Author(s):  
Christian Christensen ◽  
Sandra Mjoll Jonsdottir-Buch ◽  
Olafur Eysteinn Sigurjonsson
Keyword(s):  
Cell Culture ◽  
Growth Factors ◽  
Blood Donation ◽  
Human Platelet ◽  
Mammalian Cell Culture ◽  
Pathogen Inactivation ◽  
Platelet Concentrates ◽  
Production Methods ◽  
Lineage Differentiation

AbstractBackgroundClinical application of mesenchymal stromal cells (MSCs) usually requires an in vitro expansion step to reach clinically relevant numbers. In vitro cell expansion necessitates supplementation of basal mammalian cell culture medium with growth factors. To avoid using supplements containing animal substances, human platelet lysates (hPL) produced from expired and pathogen inactivated platelet concentrates can be used in place of fetal bovine serum. Due to lack of experience and global diversity in bacterial detection strategies, most transfusion units are currently not pathogen inactivated. As blood banks are the sole source of platelet concentrates for hPL production, it is important to ensure product safety and standardized production methods. To achieve these aims, we assessed the quality of hPL produced from expired platelet concentrates with pathogen inactivation applied after platelet lysis, as well as its ability to support MSC proliferation and tri-lineage differentiation.Methodology/principal findingsBone marrow-derived MSCs (BM-MSCs) were expanded and differentiated using hPL derived from pathogen inactivated platelet lysates (hPL-PIPL), with pathogen inactivation applied after lysis of expired platelets. Results were compared to those using hPL produced from conventional expired pathogen inactivated human platelet concentrates (hPL-PIPC), with pathogen inactivation applied after soon after blood donation. hPL-PIPL treatment had lower concentrations of soluble growth factors and cytokines than hPL-PIPC treatment. When used as supplementation in cell culture, BM-MSCs proliferated at a reduced rate, but more consistently, in hPL-PIPL than in hPL-PIPC. The ability to support tri-lineage differentiation was comparable between lysates.Conclusion/significanceThese results suggest that functional hPL can be produced from expired and untreated platelet lysates by applying pathogen inactivation after platelet lysis. When carried out post-expiration, pathogen inactivation can provide a valuable tool to further standardize global hPL production methods, increase the pool of starting material, and meet the future demand for animal-free supplements in human cell culturing.

Pharmacokinetic and toxicology assessment of INTERCEPT (S-59 and UVA treated) platelets

2001 ◽  
Vol 20 (10) ◽  
pp. 533-550 ◽  
Author(s):  
V Ciaravino ◽  
T McCullough ◽  
A D Dayan
Keyword(s):  
Ex Vivo ◽  
Reproductive Toxicity ◽  
In Vivo Studies ◽  
Pathogen Inactivation ◽  
Platelet Concentrates ◽  
Baxter Healthcare ◽  
Safety Margins ◽  
Toxicology Assessment

The pathogen inactivation process developed by Cerus and Baxter Healthcare Corporations uses the psoralen, S-59 (amotosalen) in an ex vivo photochemical treatment (PCT) process to inactivate viruses, bacteria, protozoans, and leukocytes in platelet concentrates and plasma. Studies were performed by intravenous infusion of S-59 PCT formulations-compound adsorption device (CAD) treatment and with non-UVA illuminated S-59, using doses that were multiples of potential clinical exposures. The studies comprised full pharmacokinetic, single and repeated-dose (up to 13 weeks duration) toxicity, safety pharmacology (CNS, renal, and cardiovascular), reproductive toxicity, genotoxicity, carcinogenicity testing in the p53- mouse, vein irritation, and phototoxicity. No specific target organ toxicity (clinical or histopathological), reproductive toxicity, or carcinogenicity was observed. S-59 and/or PCT formulations demonstrated CNS, ECG, and phototoxicity only at supraclinical doses. Based on the extremely large safety margins (>30,000 fold expected clinical exposures), the CNS and ECG observations are not considered to have any toxicological relevance. Additionally, after a complete assessment, mutagenicity and phototoxicity results are not considered relevant for the proposed use of INTERCEPT platelets. Thus, the results of an extensive series of in vitro and in vivo studies have not demonstrated any toxicologically relevant effects of platelet concentrates prepared by the INTERCEPT system.

scholarly journals Differentiation of Neural Stem Cells Into Cells of Oligodendroglial Lineage

2007 ◽  
Vol 50 (1) ◽  
pp. 35-41
Author(s):  
Jaroslav Mokrý ◽  
Jana Karbanová ◽  
Dana Čížková ◽  
Jan Pazour ◽  
Stanislav Filip ◽  
...  
Keyword(s):  
Stem Cells ◽  
Growth Factors ◽  
Cell Differentiation ◽  
Neural Stem Cells ◽  
Developmental Stages ◽  
Developmental Studies ◽  
Differentiation Markers ◽  
Lineage Differentiation ◽  
Massive Cell Death

We described three different conditions that induce differentiation of dissociated neural stem cells derived from mouse embryonic CNS. In the first set of experiments, where the cell differentiation was triggered by cell adhesion, removal of growth factors and serum-supplemented medium, only sporadic neuronal and astroglial cells survived longer than two weeks and the latter formed a monolayer. When differentiation was induced in serum-free medium supplemented with retinoic acid, rapid and massive cell death occurred. A prolonged survival was observed in cultivation medium supplemented with serum and growth factors EGF plus FGF-2. One third of the cells did not express cell differentiation markers and were responsible for an increase in cell numbers. The remaining cells differentiated and formed the astrocytic monolayer on which occasional neuronal cells grew. One third of the differentiated phenotypes were represented by cells of oligodendroglial lineage. Differentiation of oligodendroglial cells occurred in a stepwise mechanism because the culture contained all successive developmental stages, including oligodendrocyte progenitors, preoligodendrocytes and immature and mature oligodendrocytes. Maturing oligodendrocytes displayed immunocytochemical and morphological features characteristic of cells that undergo physiological development. The cultivation conditions that supported growth and differentiation of neural stem cells were optimal for in vitro developmental studies and the production of oligodendroglial cells.

IN VITRO CULTURE OF WHEAT TISSUES. I. CALLUS FORMATION, ORGAN REDIFFERENTIATION AND SINGLE CELL CULTURE

1969 ◽  
Vol 11 (2) ◽  
pp. 294-304 ◽  
Author(s):  
T. Shimada ◽  
T. Sasakuma ◽  
K. Tsunewaki
Keyword(s):  
Cell Culture ◽  
Growth Factors ◽  
Single Cell ◽  
Common Wheat ◽  
Callus Formation ◽  
Casein Hydrolysate ◽  
Callus Growth ◽  
Callus Cell ◽  
Basal Media

Callus induction, organ formation from callus and single callus cell culture have been tried in wheat. Though kinetin showed no effects, supplements of 2,4-D (1~10mg/1) or IAA (50mg/1) to the basal media induced calluses from seedling roots of einkorn, emmer and common wheats, and from stem pieces of common wheat. The best callus growth was obtained when casein hydrolysate (1g/1) or coconut milk (1%) was added to the basal media. Callus growth was also vigorous when 2,4-D (0.5~2.0mg/1) was added. Root formation from callus took place in all kinds of tested media, except those containing no growth factors or supplemented with 2,4-D at high concentrations (1~5mg/1). Shoot formation occurred in six cases and no growth factors were found to be specifically effective on shoot differentiation. Two plants were restored and reached maturity. Calluses of common wheat consisted of eudiploid and aneuploid cells at almost the same frequencies. The great majority of aneuploid cells had 42 ± 3 chromosomes. The restored plants showed normal chromosome constitution. Single callus-cell suspensions were obtained by the liquid culture of seeds in a shaker. A filtrate of the single cell suspension was plated on a solid agar medium, and some colonies were formed. However, plating efficiency was very low and colony growth was slow.

scholarly journals Human Platelet Lysate for Good Manufacturing Practice-Compliant Cell Production

2021 ◽  
Vol 22 (10) ◽  
pp. 5178
Author(s):  
Michaela Oeller ◽  
Sandra Laner-Plamberger ◽  
Linda Krisch ◽  
Eva Rohde ◽  
Dirk Strunk ◽  
...  
Keyword(s):  
Cell Culture ◽  
Blood Donation ◽  
Human Platelet ◽  
Raw Materials ◽  
Cell Types ◽  
Platelet Lysate ◽  
Raw Material ◽  
Specific Cell ◽  
Regulatory Requirements ◽  
Human Platelet Lysate

Numerous cell-based therapeutics are currently being tested in clinical trials. Human platelet lysate (HPL) is a valuable alternative to fetal bovine serum as a cell culture medium supplement for a variety of different cell types. HPL as a raw material permits animal serum-free cell propagation with highly efficient stimulation of cell proliferation, enabling humanized manufacturing of cell therapeutics within a reasonable timeframe. Providers of HPL have to consider dedicated quality issues regarding identity, purity, potency, traceability and safety. Release criteria have to be defined, characterizing the suitability of HPL batches for the support of a specific cell culture. Fresh or expired platelet concentrates from healthy blood donors are the starting material for HPL preparation, according to regulatory requirements. Pooling of individual platelet lysate units into one HPL batch can balance donor variation with regard to essential platelet-derived growth factors and cytokines. The increasingly applied pathogen reduction technologies will further increase HPL safety. In this review article, aspects and regulatory requirements of whole blood donation and details of human platelet lysate manufacturing are presented. International guidelines for raw materials are discussed, and defined quality controls, as well as release criteria for safe and GMP-compliant HPL production, are summarized.

scholarly journals Mechanical, chemical, structural analysis and comparative release of PDGF-AA from L-PRF, A-PRF and T-PRF - an in vitro study

2020 ◽  
Vol 24 (1) ◽  
Author(s):  
Shravanthy Ravi ◽  
Muthukumar Santhanakrishnan
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Sustained Release ◽  
Release Profile ◽  
Periodontal Therapy ◽  
Platelet Concentrates ◽  
Time Points ◽  
Regenerative Periodontal Therapy ◽  
Degradation Test

Abstract Background Platelet concentrates have been popularly used in regenerative periodontal therapy as they are autologous in origin and they provide a supernatural concentration of platelets, growth factors and leukocytes. The release profile of various growth factors is considered important during the various phases of wound healing with the most important being the inflammatory phase where the release of the growth factors help in recruitment of cells and in collagen production. With the more recent modifications of PRF namely A-PRF and T-PRF, the mechanical and chemical degradation properties have also improved. The aim of the present study was to correlate the release profile of PDGF-AA from various forms of platelet concentrates (L-PRF, A-PRF, T-PRF) based on their mechanical and chemical properties. Methods Blood samples were drawn from 2 male and 3 female systemically healthy patients between 20 and 25 years of age who were about to undergo periodontal regeneration for PRF preparation. The blood sample was immediately centrifuged using a table top centrifuge (Remi R4C) at 1060 rpm (208 x g) for 14 min for A-PRF preparation, 1960 rpm (708 x g) for 12 min for L-PRF preparation and 1960 rpm (708 x g) for 12 min in titanium tubes for T-PRF preparation. Tensile test was performed using universal testing machine. The in vitro degradation test of the prepared PRF membranes were conducted by placing the PRF membrane in 10 ml of pH 7.4 PBS on an orbital shaker set at 50 rpm. SEM evaluation of the PRF membrane was done under both low and high magnification. In order to determine the amount of released growth factor PDGF-AA at 15 min, 60 min, 8 h, 1 day, 3 days, and 10 days, samples were placed into a shaking incubator at 37 °C to allow for growth factor release into the culture media. Results On comparing the three PRF membranes, it was found that T-PRF contained the maximum tensile strength (404.61 ± 5.92 MPa) and modulus of elasticity (151.9 ± 6.92 MPa). Statistically significant differences between the three groups were found on comparing the groups for their mechanical properties. In the degradation test, it was found that the maximum amount of degradation was found in L-PRF (85.75%), followed by A-PRF (84.18%) and the least was found in T-PRF (82.27%). T-PRF released the highest amount of PDGF-AA (6060.4 pg/ml) at early time points when compared to A-PRF (5935.3 pg/ml). While T-PRF had rapid release of PDGF-AA, A-PRF had a sustained release of growth factors released at later time points. Conclusion Results from the present study indicate that A-PRF is the most favourable form of platelet concentrate in regenerative periodontal therapy as it has a sustained release of growth factors over time.

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