Acoustophoretic purification of platelets: feasibility and impact on platelet activation and function
Purity, limited platelet activation, and preservation of platelet function are important stakes of preparation of platelet concentrates (PC) for clinical use. In fact, contaminating red blood cells and leukocytes, as well as activated and/or poorly functional platelets in PC, represents a risk of po...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
2019-02-01
|
| Series: | Platelets |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/09537104.2017.1386296 |
| _version_ | 1827816918042542080 |
|---|---|
| author | Pierre Bohec Jérémie Gachelin Véronique Ollivier Thibaut Mutin Xavier Télot Benoît Ho-Tin-Noé Sandra Sanfilippo |
| author_facet | Pierre Bohec Jérémie Gachelin Véronique Ollivier Thibaut Mutin Xavier Télot Benoît Ho-Tin-Noé Sandra Sanfilippo |
| author_sort | Pierre Bohec |
| collection | DOAJ |
| description | Purity, limited platelet activation, and preservation of platelet function are important stakes of preparation of platelet concentrates (PC) for clinical use. In fact, contaminating red blood cells and leukocytes, as well as activated and/or poorly functional platelets in PC, represents a risk of poor efficiency and adverse side effects during platelet transfusion. Therefore, optimization of preparation and storage of PC is still an active field of research. Shear-induced platelet activation is an unwanted side effect of the hard-spin (up to 5000g) step of centrifugation-based methods currently used in blood banks to prepare PC from whole blood samples. Here, we evaluated the effectiveness of an acoustic-based fractionation device for the isolation of human platelets from whole blood bags. The purity, activation status, and functionality of platelets isolated by acoustopheresis were compared with those of platelets isolated using a reference protocol known to produce limited platelet activation and consisting of two consecutive soft-spin centrifugations (120g and 1200g). Platelet concentration and purity were determined using an automated hematology analyzer. Platelet activation status and platelet reactivity to collagen and thrombin were assessed in flow cytometry by measurement of surface expression of P-selectin and activated integrin αIIbβ3. The ability of isolated platelets to incorporate into a thrombus when transfused to NOD/SCID mice was investigated by intravital microscopy using the ferric chloride-induced thrombosis model. Blood fractionation by acoustophoresis led to the elimination of more than 80% of red blood cells and leukocytes from the platelet fraction, whose mean purity was of 92.8 ± 12.8%. The activation status and reactivity to collagen and thrombin of acoustophoresis-isolated platelets were similar to those of platelets isolated by soft-spin centrifugation. Finally, acoustophoresis-isolated platelets were tethered, adhered to the vessel wall, and incorporated into a growing thrombus following ferric chloride-induced vascular injury. Together, our results indicate that acoustophoresis is a suitable method for the isolation of human platelets with minimal platelet activation and preservation of platelet function. |
| first_indexed | 2024-03-12T00:27:06Z |
| format | Article |
| id | doaj.art-771cd35f2f9c45d8a47178174a37d897 |
| institution | Directory Open Access Journal |
| issn | 0953-7104 1369-1635 |
| language | English |
| last_indexed | 2024-03-12T00:27:06Z |
| publishDate | 2019-02-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Platelets |
| spelling | doaj.art-771cd35f2f9c45d8a47178174a37d8972023-09-15T10:31:58ZengTaylor & Francis GroupPlatelets0953-71041369-16352019-02-0130217418010.1080/09537104.2017.13862961386296Acoustophoretic purification of platelets: feasibility and impact on platelet activation and functionPierre Bohec0Jérémie Gachelin1Véronique Ollivier2Thibaut Mutin3Xavier Télot4Benoît Ho-Tin-Noé5Sandra Sanfilippo6Hôpital Saint-LouisHôpital Saint-LouisHôpital BichatHôpital Saint-LouisHôpital Saint-LouisHôpital BichatHôpital Saint-LouisPurity, limited platelet activation, and preservation of platelet function are important stakes of preparation of platelet concentrates (PC) for clinical use. In fact, contaminating red blood cells and leukocytes, as well as activated and/or poorly functional platelets in PC, represents a risk of poor efficiency and adverse side effects during platelet transfusion. Therefore, optimization of preparation and storage of PC is still an active field of research. Shear-induced platelet activation is an unwanted side effect of the hard-spin (up to 5000g) step of centrifugation-based methods currently used in blood banks to prepare PC from whole blood samples. Here, we evaluated the effectiveness of an acoustic-based fractionation device for the isolation of human platelets from whole blood bags. The purity, activation status, and functionality of platelets isolated by acoustopheresis were compared with those of platelets isolated using a reference protocol known to produce limited platelet activation and consisting of two consecutive soft-spin centrifugations (120g and 1200g). Platelet concentration and purity were determined using an automated hematology analyzer. Platelet activation status and platelet reactivity to collagen and thrombin were assessed in flow cytometry by measurement of surface expression of P-selectin and activated integrin αIIbβ3. The ability of isolated platelets to incorporate into a thrombus when transfused to NOD/SCID mice was investigated by intravital microscopy using the ferric chloride-induced thrombosis model. Blood fractionation by acoustophoresis led to the elimination of more than 80% of red blood cells and leukocytes from the platelet fraction, whose mean purity was of 92.8 ± 12.8%. The activation status and reactivity to collagen and thrombin of acoustophoresis-isolated platelets were similar to those of platelets isolated by soft-spin centrifugation. Finally, acoustophoresis-isolated platelets were tethered, adhered to the vessel wall, and incorporated into a growing thrombus following ferric chloride-induced vascular injury. Together, our results indicate that acoustophoresis is a suitable method for the isolation of human platelets with minimal platelet activation and preservation of platelet function.http://dx.doi.org/10.1080/09537104.2017.1386296acoustophoresishuman blood fractionationplatelet activation |
| spellingShingle | Pierre Bohec Jérémie Gachelin Véronique Ollivier Thibaut Mutin Xavier Télot Benoît Ho-Tin-Noé Sandra Sanfilippo Acoustophoretic purification of platelets: feasibility and impact on platelet activation and function Platelets acoustophoresis human blood fractionation platelet activation |
| title | Acoustophoretic purification of platelets: feasibility and impact on platelet activation and function |
| title_full | Acoustophoretic purification of platelets: feasibility and impact on platelet activation and function |
| title_fullStr | Acoustophoretic purification of platelets: feasibility and impact on platelet activation and function |
| title_full_unstemmed | Acoustophoretic purification of platelets: feasibility and impact on platelet activation and function |
| title_short | Acoustophoretic purification of platelets: feasibility and impact on platelet activation and function |
| title_sort | acoustophoretic purification of platelets feasibility and impact on platelet activation and function |
| topic | acoustophoresis human blood fractionation platelet activation |
| url | http://dx.doi.org/10.1080/09537104.2017.1386296 |
| work_keys_str_mv | AT pierrebohec acoustophoreticpurificationofplateletsfeasibilityandimpactonplateletactivationandfunction AT jeremiegachelin acoustophoreticpurificationofplateletsfeasibilityandimpactonplateletactivationandfunction AT veroniqueollivier acoustophoreticpurificationofplateletsfeasibilityandimpactonplateletactivationandfunction AT thibautmutin acoustophoreticpurificationofplateletsfeasibilityandimpactonplateletactivationandfunction AT xaviertelot acoustophoreticpurificationofplateletsfeasibilityandimpactonplateletactivationandfunction AT benoithotinnoe acoustophoreticpurificationofplateletsfeasibilityandimpactonplateletactivationandfunction AT sandrasanfilippo acoustophoreticpurificationofplateletsfeasibilityandimpactonplateletactivationandfunction |