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 -