Table 4 Protocols used for the production of HPL
References | Starting material | Donor number | Preliminary steps | Dilution | Platelet lysis | Debris removal | Fibrin removal | Insoluble protein removal | Filtration step(s) | Storage | Further steps | Heparin addition | Standardization and characterization |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Shanbhag et al. [11] | Whole blood-derived Platelet concentrate | Four different PCs (each PC containing buffy coats from five donors) | – | – | Multiple freeze/thaw cycles | Centrifugation 3000 g, 15 min at 4 °C | – | – | – | − 80 °C | – | 1 IU/mL | 1) > 2 × 1011 platelets 2) virological test |
Fuocoet al. [16] | Platelet concentrate | 4 | – | – | 4 freeze/thaw cycles | 4 × centrifugation 3600 g, 30 min | – | – | 0.22 µm filter | − 20 °C | – | 2 U/mL | – |
Palombella et al. [5] | Commercial: HPL FD, GMP Grade (Antibodies-online.com) | 150–300 | nd | – | 3 freeze/thaw cycles | yes | yes | nd | nd | nd | nd | – | – |
Gao et al. [17] | Platelet concentrate | Minimum 15 donors | – | – | 3 freeze (− 80 °C)/thaw (37 °C) cycles | Centrifugation 3000 g, 30 min at 20 °C | Incubation with 23 mM CaCl2 and glass beads at 24–26 °C for 1 h Centrifugation at 6000 g, 30 min at 20 °C | Heat-inactivation at 56 °C, 30 min Centrifugation | 0.2 µm filter | − 20 °C | – | – | – |
Becherucci et al. [18] | Whole blood | 40 | Centrifugation to obtain buffy coat CompoStop Flexible kit to obtain platelet concentrate (leukocyte depletion by pressure filtration) Centrifugation 400 g, 9 min at 22 °C Centrifugation 457 g, 30 min at 4 °C Filtration | Thawed fresh-frozen AB group plasma | 3 freeze/thaw cycles | 2 × centrifugation 4579 g, 10 min at 20 °C | – | – | 0.45 µm filter 0.2 µm filter | − 80 °C | – | 40 U/mL | 1.5 × 109–2.4 × 109 platelets/mL (before lysis) Sterility test Endotoxin test |
Boraldi et al. [19] | Commercial: Stemulate, cook medical (+ / − heparin) Macopharma Lyset (Sclavo Diagnostic International) | nd | nd | nd | nd | nd | nd | nd | nd | nd | nd | 1 U/mL | nd |
Pierce et al. [20] | PRP (apheresis, frozen, expired ≤ 3 d) | 49–109 | – | – | Thawing (4 °C) | PL-P: Centrifugation 4000 g for 20 min | PL-S: Incubation with 20% (w/v) CaCl2 at 4 °C for 24 h Centrifugation 4000 g for 20 min | – | Details not described (proprietary process) | – | – | PL-P: 2 U/mL | Sterility test Biochemical analyses Functional assessment |
Fernandez-Rebollo et al. [21] | Platelet apheresis product | At least 5 | – | – | 2 freeze (− 80 °C)/thaw cycles (37 °C) | Centrifugation 2600 g for 30 min | – | – | 0.2 µm filter | − 80 °C | – | 0.61 U | – |
Frese et al. [22] | Platelet apheresis product | 12 | – | – | Thawing (37 °C) | Centrifugation 5000 rpm for 30 min | – | – | 0.22 µm filter | − 80 °C | Centrifugation 5000 rpm for 30 min before use | 1 U/mL | – |
Juhl et al. [23] | Commercial: PLTMax Stemulate PL-S (heparin-required) Stemulate PL-SP (heparin-free) | PLTMax: Multiple donors Stemulate: Multiple donors | nd | nd | nd | nd | nd | nd | nd | nd | nd | 10 U | PLTMax: Sterility, endotoxin, mycoplasma test BMSC growth assay Stemulate: ISO9001:2015 compliant Sterility, endotoxin, mycoplasma test BMSC growth assay |
Riis et al. [20) | Commercial: Stemulate, Cook Medical | Multiple donors | nd | nd | nd | nd | nd | nd | nd | nd | nd | nd | ISO9001:2015 compliant Sterility, endotoxin, mycoplasma test BMSC growth assay |
Castrèn et al. [25] | Platelet apheresis product | 4 | centrifugation | AB plasma | 5 freeze/thaw cycles | – | – | – |  | – | – | 40 U/mL | 300 × 109 platelets/each platelet unit 0.8 × 108 platelets/mL final concentration BMSC growth assay |
Hildner et al. [26] | Whole blood | 36 | Centrifugation 3939 g for 11 min to isolate buffy coat 2 h at 22 °C Centrifugation 404 g for 6 min Manual squeezing of the bag in plasma extractor to obtain PRP | – | Thawing (37 °C) | Centrifugation 2000 g for 10 min | – | – | 0.22 µm filter | − 80 °C | – | 2 U/mL | 1–2 × 109 platelets/mL (before lysis) Quantification of TGFβ1, bFGF, IGF-1, PDGF-BB |
Muraglia et al. [27] | Buffy coats | 300–400 | Centrifugation 1100 rpm for 10 min to obtain PRP Centrifugation 2600 rpm for 20 min | Platelet Poor Plasma | 3 freeze/thaw cycles | Centrifugation at high speed for 20 min at RT | – | – | – | − 20 °C | Lyophilization and restoration with sterile water before use | 40 U/mL | 10 × 109 platelets/mL (before lysis) Endotoxin, mycoplasma test Quantification of PDGF-BB, VEGF, fibrinogen, hemoglobin |
Castiglia et al. [28] | Whole blood | 60 | Centrifugation to isolate buffy coat Automatically separated through a leukoreduction filter (TACSI system) | AB-group plasma | 3 freeze (− 35 °C)/thaw (37 °C) cycles | Centrifugation 5000 g for 8 min | – | – | – | − 35 °C | – | 20 U/mL | Virological test |
Fekete et al. [13] | Whole blood | 12 | 3–22 h freezing Centrifugation to obtain buffy coat Irradiation with 30 Gy Quarantine storage − 30 °C for up to 18 months | – | Thawing at 37 °C and store at 4 °C | Centrifugation 4000 rpm for 10 min at 20 °C | – | – | 0.8/0.45 μm filter 0.45/0.2 or 0.35/0.2 μm filter |  − 80 °C to − 30 °C | 1200 × g for 10 min 50,000 × g for 60 min | 2 U/mL | At least 109 platelets/mL Sterility test Endotoxin test |
Bernardi et al. [29] | PRP | 4–6 | – | – | Sonication by ultrasound stimulation at 20 kHz for 30 min OR 3 freeze (− 80 °C)/thaw cycles (37 °C) | Centrifugation 1600 g for 15 min at RT | – | – | 70 µm cell Strainer | − 20 °C | – | – | Quantification of PDGF-AB |
Kinzebach et al. [12] | Buffy coat | 8 | – | AB plasma | 1 freeze (− 30 °C)/thaw (37 °C) cycle | Centrifugation 2000 g for 20 min at RT | – | – | 0.45 µm filter (complete medium) | − 30 °C | Centrifugation 2000 g for 10 min | 5 U/mL | – |
Menard et al. [30] | Buffy coat | 4 | storage at 22 °C for no longer than 5 days | – | Freeze (− 30 °C)/thaw 37 °C | Centrifugation 4000 rpm 10 min at 20 °C |  |  |  |  |  | 2 U/mL | Sterility test Endotoxin test |
Trojahn Kølle et al. [31] | Buffy coat | 40 | Dilution and freeze (− 40 °C) | AB plasma | Thawing (37 °C) | Centrifugation 4000 g for 15 min | – | – | – | − 80 °C | – | 2 U/mL | – |
Warnke et al. [32] | Platelet apheresis product | 10 | Freeze (− 80 °C) | – | Thawing (37 °C for 10 min) | Centrifugation 4000 g for 15 min at RT | – | – | – | − 80 °C | – | 2 U/mL | – |
Azouna et al. [33] | Whole blood | at least 10 | Incubation at 22 °C for 16 h Centrifugation 4250 g for 13 min at 22 °C to obtain buffy coat Centrifugation 341 g for 6 min at 22 °C White blood cell depletion Freeze (− 30 °C) | AB plasma | Thawing (37 °C) | Centrifugation 1400 g for 20 min | – | – | 0.22 µm filter | − 80 °C | Centrifugation 4000 g for 15 min | – | Platelet concentration White/red blood cell contamination Sterility test |
Gottipamula et al. [34] | Platelet concentrate | 30 | – | – | 5 freeze/thaw cycles | Centrifugation 4000 g for 15 min | – | – | – | – | – | – | – |
Cholewa et al. [35] | Platelet concentrate | 5 | Supplementation with acid-citrate-dextrose (1:1, v/v) | – | 2 freeze (− 80 °C)/thaw (37 °C) cycles | Centrifugation 2600 g for 30 min at 4 °C | – | – | 0.22 µm filter | − 80 °C | – | 2 U/mL | 1–2.1 × 109 platelets/mL (average in platelet units) |
Flemming et al. [36] | Whole blood | 40 | Isolation of buffy coat Centrifugation 340 g for 6 min at 22 °C Filtration for leukocyte depletion | AB plasma | 1 freeze (− 30 °C)/thaw (37 °C) | – | – | – | – | − 30 °C | Centrifugation 4000 g for 15 min at RT | 2 U/mL | – |
Castegnaro et al. [37] | Pooled AB plasma | 5 | Centrifugation 240 g for 10 min | – | Freeze and sonication | Centrifugation 1500 g for 30 min | – | – | – | − 20 °C | – | – | 0.686 × 109 platelets/mL (average in pooled plasma) |
Chevallier et al. [38] | Platelet apheresis product | 4 | – | – | Freeze − 80 °C | Centrifugation 1400 g | – | – | – | – | – | 2 U/mL |  > 1 × 109 platelets/mL (in initial apheresis product) |
Horn et al. [14] | Platelet concentrate | autologous | Supplementation with acid-citrate-dextrose (1:1, v/v) | – | 2 freeze (− 80 °C)/thaw (37 °C) cycles | Centrifugation 2600 g for 30 min | – | – | 0.2 µm filter | − 80 °C | 2 U/mL | 2 U/mL | 1–2.1 × 109 platelets/mL (average in platelet concentrates) |
Schallmoser et al. [39] | Whole blood | at least 40 | Incubation at 22 °C for 16 h Centrifugation 4250 g for 13 min at 22 °C to obtain buffy coat Centrifugation 341 g for 6 min at 22 °C White blood cell depletion Freeze (− 30 °C) | AB plasma | Thawing (37 °C) | Centrifugation 1400 g for 20 min | – | – | – | − 80 °C | – | 2 U/mL | – |
Bieback et al. [40] | Buffy coat | 40–50 | Centrifugation 340 g for 6 min at 22 °C Inline filtration for leukocyte depletion Freeze (− 30 °C) | AB plasma | Thawing (37 °C) | Centrifugation 1400 g for 20 min | – | – | – | − 30 °C | Centrifugation 4000 g for 15 min | 2 U/mL | – |
Blande et al. [15] | Whole blood | autologous | – | – | 4 freeze (− 80 °C)/thaw (37 °C) cycles | 4 × Centrifugation 3313 g for 30 min | – | – | 0.22 µm filter | − 20 °C | – | 2 U/mL | 0.91 × 109 platelet/mL (average in whole blood) |
Prins et al. [41] | Platelet apheresis product | 5 | – | – | Freeze − 80 °C | – | – | – | – | – | Centrifugation 750 g for 10 min | 10 U/mL | 1 × 109 platelet/mL (average in platelet products) |
Capelli et al. [42] | Whole blood | 1 | Light-spin centrifugation to isolate PRP Heavy-spin centrifugation to concentrate platelets | – | Freeze − 40 °C | 3000 rpm for 10 min | – | – | – | − 20 °C | – | 1000 U | 1.2 ± 0.4 × 109 platelets/mL (average in platelet concentrate) |
Schallmoser et al. [43] | Whole blood | 40 | Incubation at 22 °C for 16 h Centrifugation 4250 g for 13 min at 22 °C to obtain buffy coat Centrifugation 341 g for 6 min at 22 °C White blood cell depletion Freeze (− 30 °C) | AB plasma | Thawing (37 °C) | Centrifugation 1400 g for 20 min | – | – | 0.22 µm filter | − 30 °C | – | 2 U/mL | Platelet concentration White and red blood cell contamination Sterility test |
Doucet et al. [3] | Platelet apheresis product | 10 | – | – | Freeze − 80 °C | Centrifugation 900 g | – | – | – | – | – | 2 U/mL | 1 × 109 platelets/mL (in initial apheresis product) |