Details
| Original language | English |
|---|---|
| Article number | 7654 |
| Pages (from-to) | 1-19 |
| Number of pages | 19 |
| Journal | International Journal of Molecular Sciences |
| Volume | 21 |
| Issue number | 20 |
| Publication status | Published - 16 Oct 2020 |
Abstract
Donor platelet transfusion is currently the only efficient treatment of life-threatening thrombocytopenia, but it is highly challenged by immunological, quality, and contamination issues, as well as short shelf life of the donor material. Ex vivo produced megakaryocytes and platelets represent a promising alternative strategy to the conventional platelet transfusion. However, practical implementation of such strategy demands availability of reliable biobanking techniques, which would permit eliminating continuous cell culture maintenance, ensure time for quality testing, enable stock management and logistics, as well as availability in a ready-to-use manner. At the same time, protocols applying DMSO-based cryopreservation media were associated with increased risks of adverse long-term side effects after patient use. Here, we show the possibility to develop cryopreservation techniques for iPSC-derived megakaryocytes under defined xeno-free conditions with significant reduction or complete elimination of DMSO. Comprehensive phenotypic and functional in vitro characterization of megakaryocytes has been performed before and after cryopreservation. Megakaryocytes cryopreserved DMSO-free, or using low DMSO concentrations, showed the capability to produce platelets in vivo after transfusion in a mouse model. These findings propose biobanking approaches essential for development of megakaryocyte-based replacement and regenerative therapies.
Keywords
- Biobanking, Cytotoxicity, Dimethyl sulfoxide, Ethylene glycol, Induced pluripotent stem cells (iPSC), Megakaryocytes, Mouse model, Platelets, Propane-1,2-diol, Transfusion
ASJC Scopus subject areas
- Chemical Engineering(all)
- Catalysis
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Chemistry(all)
- Spectroscopy
- Computer Science(all)
- Computer Science Applications
- Chemistry(all)
- Physical and Theoretical Chemistry
- Chemistry(all)
- Organic Chemistry
- Chemistry(all)
- Inorganic Chemistry
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In: International Journal of Molecular Sciences, Vol. 21, No. 20, 7654, 16.10.2020, p. 1-19.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Towards Reduction or Substitution of Cytotoxic DMSO in Biobanking of Functional Bioengineered Megakaryocytes
AU - Pogozhykh, Denys
AU - Eicke, Dorothee
AU - Gryshkov, Oleksandr
AU - Wolkers, Willem F.
AU - Schulze, Kai
AU - Guzmán, Carlos A.
AU - Blasczyk, Rainer
AU - Figueiredo, Constança
N1 - Funding information: This study received support from the Erich und Gertrud Roggenbuck Stiftung.
PY - 2020/10/16
Y1 - 2020/10/16
N2 - Donor platelet transfusion is currently the only efficient treatment of life-threatening thrombocytopenia, but it is highly challenged by immunological, quality, and contamination issues, as well as short shelf life of the donor material. Ex vivo produced megakaryocytes and platelets represent a promising alternative strategy to the conventional platelet transfusion. However, practical implementation of such strategy demands availability of reliable biobanking techniques, which would permit eliminating continuous cell culture maintenance, ensure time for quality testing, enable stock management and logistics, as well as availability in a ready-to-use manner. At the same time, protocols applying DMSO-based cryopreservation media were associated with increased risks of adverse long-term side effects after patient use. Here, we show the possibility to develop cryopreservation techniques for iPSC-derived megakaryocytes under defined xeno-free conditions with significant reduction or complete elimination of DMSO. Comprehensive phenotypic and functional in vitro characterization of megakaryocytes has been performed before and after cryopreservation. Megakaryocytes cryopreserved DMSO-free, or using low DMSO concentrations, showed the capability to produce platelets in vivo after transfusion in a mouse model. These findings propose biobanking approaches essential for development of megakaryocyte-based replacement and regenerative therapies.
AB - Donor platelet transfusion is currently the only efficient treatment of life-threatening thrombocytopenia, but it is highly challenged by immunological, quality, and contamination issues, as well as short shelf life of the donor material. Ex vivo produced megakaryocytes and platelets represent a promising alternative strategy to the conventional platelet transfusion. However, practical implementation of such strategy demands availability of reliable biobanking techniques, which would permit eliminating continuous cell culture maintenance, ensure time for quality testing, enable stock management and logistics, as well as availability in a ready-to-use manner. At the same time, protocols applying DMSO-based cryopreservation media were associated with increased risks of adverse long-term side effects after patient use. Here, we show the possibility to develop cryopreservation techniques for iPSC-derived megakaryocytes under defined xeno-free conditions with significant reduction or complete elimination of DMSO. Comprehensive phenotypic and functional in vitro characterization of megakaryocytes has been performed before and after cryopreservation. Megakaryocytes cryopreserved DMSO-free, or using low DMSO concentrations, showed the capability to produce platelets in vivo after transfusion in a mouse model. These findings propose biobanking approaches essential for development of megakaryocyte-based replacement and regenerative therapies.
KW - Biobanking
KW - Cytotoxicity
KW - Dimethyl sulfoxide
KW - Ethylene glycol
KW - Induced pluripotent stem cells (iPSC)
KW - Megakaryocytes
KW - Mouse model
KW - Platelets
KW - Propane-1,2-diol
KW - Transfusion
UR - http://www.scopus.com/inward/record.url?scp=85093942993&partnerID=8YFLogxK
U2 - 10.3390/ijms21207654
DO - 10.3390/ijms21207654
M3 - Article
C2 - 33081128
AN - SCOPUS:85093942993
VL - 21
SP - 1
EP - 19
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
SN - 1661-6596
IS - 20
M1 - 7654
ER -