In vitro and ex vivo platelet-producing systems based on their design
| Publication . | Brief description of system . | Platelet generated per input cell . | MK species and origin . | Platelet functionality . | Basis for design . |
|---|---|---|---|---|---|
| Choi et al44 | Static cell culture | 240 per (proplatelet extending) MK | Human peripheral blood | Platelets aggregated in response to thrombin and ADP/fibrinogen. CD62 and activated integrin αIIbβ3 expression increased in response to thrombin and ADP, respectively. | Static or perfused cellular culture |
| Matsunaga et al78 | Multistage cell culture | 4 × 104 per starting CD34+ cell | Human umbilical cord blood | Platelets aggregated in response to ADP/fibrinogen. Platelet CD62P and activated integrin αIIbβ3 expression increased in response to ADP. | |
| Takayama et al79 | Static cell culture | ∼48 per hESC∗ | hESC | Platelets increased activated integrin αIIbβ3 expression in response to thrombin and ADP. Filopodia are also formed on fibrinogen coated surface. | |
| Lu et al80 | Static cell culture | 6.7 per MK | hESC | Platelets formed microaggregates, facilitated clot formation/retraction, and formed lamellipodia and filopodia in response to thrombin activation. In vivo function documented by incorporation of produced platelet-like particles into a newly formed thrombus in mice. | |
| Sullenbarger et al81 | Perfused 3D hydrogel bioreactor | 19.6 per starting CD34+ cell | Human umbilical cord blood | Platelets showed increased CD62 and CD63 expression following maximal thrombin stimulation. No statistical analysis was shown. Preactivation of platelets was present. | |
| Lasky et al82 | Added shear forces and oxygenation | Twofold to threefold increase in platelet numbers | Platelet aggregation and CD62 and CD63 expression measured. Statistical analysis missing but a degree of aggregation occurred and an increase in the percentage of CD62- and CD63-expressing platelets when activated with thrombin. | ||
| Dunois-larde et al69 | Perfusion at high shear rates on vWf | Not stated | Human umbilical cord blood and bone marrow | Platelets spread on fibrinogen-coated surface and increased CD62P expression in response to thrombin | Shear forces; turbulent and/or laminar flow |
| Blin et al83 | PDMS microfluidics pillar forest coated with vWf | 3.7 per MK | Human umbilical cord blood | Platelets spread on fibrinogen-coated surface and exhibit cytoskeletal changes in response to thrombin. Significant increase in integrin αIIbβ3 in response to TRAP agonist. | |
| Ito et al84 | Microfluidic pillar chamber Static cell culture 8 L tank with turbulent flow | 14 per MK <20 per MK 70-80 per MK | Immortalized MK cell lines derived from hiPSCs | Extensive flow cytometric, aggregation, and in vivo mouse bleeding time testing showing functional platelets in the various iterations of the bioreactor | |
| Strassel et al85 | Successive pipetting of cultured cells | 28 per MK | Human peripheral blood | In vitro function of platelet-like particles observed in a flow chamber and in vivo function documented by the incorporation of produced platelet-like particles into a newly formed thrombus in mice | |
| Pongérard et al86 | Polypropylene spheres' fluidic system producing turbulent and laminar flow | 100 per MK | Increased expression of αIIbβ3 in response to a range of agonists. Higher survival of platelets when injected into a mouse when compared with control. | ||
| Nakagawa et al87 | Scaffolds with flow and ordered pores | 0.6 per MK∗ | hESC or hiPSCs KhES-3 hESC clone TkDA3-4 hiPSC clone Mouse C3H10T1/2 cell line | Increased expression of activated αIIbβ3 in ESC and iPSC-derived platelets in response to a range of agonists (no statistical analysis, n = 2) | Bone-marrow mimetic |
| Avanzi et al88 | Nanofiber membranes and flow | 100 per MK | Human umbilical cord blood | Increased CD62P expression upon stimulation with PMA (n = 1 shown) | |
| Martinez et al89 | Bioreactor with slits and flow | Not clearly stated | Human peripheral blood | Increased expression of CD62P on platelet-like particles upon stimulation with thrombin (no statistical analysis). Adherence to fibrinogen coated surface and spreads upon thrombin stimulation. | |
| Tozzi et al90 | Bone-marrow niche replicate synthesized in silk fibroin | 124 per MK∗ | Human umbilical cord blood | Increased expression of αIIbβ3 in response to thrombin | |
| Di Buduo et al91 | 3D porous silk sponge with ECM components, cytokines, and endothelium-derived proteins | 5.6 per (seeded) MK∗ | Human umbilical cord blood | Adhesion to collagen under flow conditions and established actin reorganization. Increased expression of CD31 and CD42b upon stimulation with multiple agonists at once. Produced platelets mixed with native platelets during an ex vivo clot assay. | |
| Pallotta et al92 | Silk microtubular bioreactor | ∼200 per (proplatelet extending) MK (7% of all MKs) | Human umbilical cord blood | Increased expression of αIIbβ3 and CD62P following thrombin stimulation | |
| Thon et al93 | PDMS microfluidics with ECM components + endothelial cells | 42 per murine MK (∼20 in static culture) ∼30 per human MK | Primary mouse bone marrow–derived MKs hiPSCs | Platelets form lamellipodia and filipodia when activated | |
| Fujiyama et al94 | Ex vivo porcine thighbone | 4.4 per MK∗ | Human umbilical cord blood | Increased expression of CD62P following thrombin stimulation (no statistical analysis) | |
| Zhao et al50 | Ex vivo murine lungs vasculature perfusion Microfluidic branching perfusion | 2997 per MK 492 per MK | Primary mouse bone marrow–derived MKs | In vitro and ex vivo system–produced platelets increased expression of αIIbβ3 and CD62P following stimulation with thrombin or CRP-XL. Ex vivo–generated platelets also incorporated into an in vitro thrombus containing native platelets. | Lung vasculature mimetic |
| Publication . | Brief description of system . | Platelet generated per input cell . | MK species and origin . | Platelet functionality . | Basis for design . |
|---|---|---|---|---|---|
| Choi et al44 | Static cell culture | 240 per (proplatelet extending) MK | Human peripheral blood | Platelets aggregated in response to thrombin and ADP/fibrinogen. CD62 and activated integrin αIIbβ3 expression increased in response to thrombin and ADP, respectively. | Static or perfused cellular culture |
| Matsunaga et al78 | Multistage cell culture | 4 × 104 per starting CD34+ cell | Human umbilical cord blood | Platelets aggregated in response to ADP/fibrinogen. Platelet CD62P and activated integrin αIIbβ3 expression increased in response to ADP. | |
| Takayama et al79 | Static cell culture | ∼48 per hESC∗ | hESC | Platelets increased activated integrin αIIbβ3 expression in response to thrombin and ADP. Filopodia are also formed on fibrinogen coated surface. | |
| Lu et al80 | Static cell culture | 6.7 per MK | hESC | Platelets formed microaggregates, facilitated clot formation/retraction, and formed lamellipodia and filopodia in response to thrombin activation. In vivo function documented by incorporation of produced platelet-like particles into a newly formed thrombus in mice. | |
| Sullenbarger et al81 | Perfused 3D hydrogel bioreactor | 19.6 per starting CD34+ cell | Human umbilical cord blood | Platelets showed increased CD62 and CD63 expression following maximal thrombin stimulation. No statistical analysis was shown. Preactivation of platelets was present. | |
| Lasky et al82 | Added shear forces and oxygenation | Twofold to threefold increase in platelet numbers | Platelet aggregation and CD62 and CD63 expression measured. Statistical analysis missing but a degree of aggregation occurred and an increase in the percentage of CD62- and CD63-expressing platelets when activated with thrombin. | ||
| Dunois-larde et al69 | Perfusion at high shear rates on vWf | Not stated | Human umbilical cord blood and bone marrow | Platelets spread on fibrinogen-coated surface and increased CD62P expression in response to thrombin | Shear forces; turbulent and/or laminar flow |
| Blin et al83 | PDMS microfluidics pillar forest coated with vWf | 3.7 per MK | Human umbilical cord blood | Platelets spread on fibrinogen-coated surface and exhibit cytoskeletal changes in response to thrombin. Significant increase in integrin αIIbβ3 in response to TRAP agonist. | |
| Ito et al84 | Microfluidic pillar chamber Static cell culture 8 L tank with turbulent flow | 14 per MK <20 per MK 70-80 per MK | Immortalized MK cell lines derived from hiPSCs | Extensive flow cytometric, aggregation, and in vivo mouse bleeding time testing showing functional platelets in the various iterations of the bioreactor | |
| Strassel et al85 | Successive pipetting of cultured cells | 28 per MK | Human peripheral blood | In vitro function of platelet-like particles observed in a flow chamber and in vivo function documented by the incorporation of produced platelet-like particles into a newly formed thrombus in mice | |
| Pongérard et al86 | Polypropylene spheres' fluidic system producing turbulent and laminar flow | 100 per MK | Increased expression of αIIbβ3 in response to a range of agonists. Higher survival of platelets when injected into a mouse when compared with control. | ||
| Nakagawa et al87 | Scaffolds with flow and ordered pores | 0.6 per MK∗ | hESC or hiPSCs KhES-3 hESC clone TkDA3-4 hiPSC clone Mouse C3H10T1/2 cell line | Increased expression of activated αIIbβ3 in ESC and iPSC-derived platelets in response to a range of agonists (no statistical analysis, n = 2) | Bone-marrow mimetic |
| Avanzi et al88 | Nanofiber membranes and flow | 100 per MK | Human umbilical cord blood | Increased CD62P expression upon stimulation with PMA (n = 1 shown) | |
| Martinez et al89 | Bioreactor with slits and flow | Not clearly stated | Human peripheral blood | Increased expression of CD62P on platelet-like particles upon stimulation with thrombin (no statistical analysis). Adherence to fibrinogen coated surface and spreads upon thrombin stimulation. | |
| Tozzi et al90 | Bone-marrow niche replicate synthesized in silk fibroin | 124 per MK∗ | Human umbilical cord blood | Increased expression of αIIbβ3 in response to thrombin | |
| Di Buduo et al91 | 3D porous silk sponge with ECM components, cytokines, and endothelium-derived proteins | 5.6 per (seeded) MK∗ | Human umbilical cord blood | Adhesion to collagen under flow conditions and established actin reorganization. Increased expression of CD31 and CD42b upon stimulation with multiple agonists at once. Produced platelets mixed with native platelets during an ex vivo clot assay. | |
| Pallotta et al92 | Silk microtubular bioreactor | ∼200 per (proplatelet extending) MK (7% of all MKs) | Human umbilical cord blood | Increased expression of αIIbβ3 and CD62P following thrombin stimulation | |
| Thon et al93 | PDMS microfluidics with ECM components + endothelial cells | 42 per murine MK (∼20 in static culture) ∼30 per human MK | Primary mouse bone marrow–derived MKs hiPSCs | Platelets form lamellipodia and filipodia when activated | |
| Fujiyama et al94 | Ex vivo porcine thighbone | 4.4 per MK∗ | Human umbilical cord blood | Increased expression of CD62P following thrombin stimulation (no statistical analysis) | |
| Zhao et al50 | Ex vivo murine lungs vasculature perfusion Microfluidic branching perfusion | 2997 per MK 492 per MK | Primary mouse bone marrow–derived MKs | In vitro and ex vivo system–produced platelets increased expression of αIIbβ3 and CD62P following stimulation with thrombin or CRP-XL. Ex vivo–generated platelets also incorporated into an in vitro thrombus containing native platelets. | Lung vasculature mimetic |
Original publication, a brief description of the design, the platelet production efficiency, the species/cell origin of the MK, and the functionality of the produced cells are included. Efficiency is described as platelets generated per input MK (or per original stem cell if MK numbers were not provided).
ADP, adenosine 5′-diphosphate; ESC, embryonic stem cells; hESC, human embryonic stem cells; PDMS, polydimethylsiloxane.
When not clearly stated in the publication, the exhibited data were analyzed to estimate the platelet-producing efficiency.