Establishing a method for the cryopreservation of viable peripheral blood mononuclear cells in the International Space Station

• Published in: NPJ microgravity Vol. 10; no. 1; pp. 84 - 9
• Main Authors: Ishii, Hiroto, Endo, Rin, Hamanaka, Sanae, Hidaka, Nobuyuki, Miyauchi, Maki, Hagiwara, Naho, Miyao, Takahisa, Yamamori, Tohru, Aiba, Tatsuya, Akiyama, Nobuko, Akiyama, Taishin
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 09-08-2024; Nature Publishing Group; Nature Portfolio
• Subjects: 631/250; 692/308/2778; 692/700; Applied Microbiology; Biomedical and Life Sciences; Biotechnology; Blood; Blood cells; Cell surface; Classical and Continuum Physics; Cryopreservation; Cryoprotectors; Dimensional analysis; Flow cytometry; Freezing; Hydroxyethyl starch; Immunology; Leukocytes (mononuclear); Life Sciences; Peripheral blood mononuclear cells; Space Exploration and Astronautics; Space Sciences (including Extraterrestrial Physics; Space stations; Surface markers; Thawing
• ISSN: 2373-8065; 2373-8065
• DOI: 10.1038/s41526-024-00423-2

The analysis of cells frozen within the International Space Station (ISS) will provide crucial insights into the impact of the space environment on cellular functions and properties. The objective of this study was to develop a method for cryopreserving blood cells under the specific constraints of the ISS. In a ground experiment, mouse blood was directly mixed with a cryoprotectant and gradually frozen at −80 °C. Thawing the frozen blood sample resulted in the successful recovery of viable mononuclear cells when using a mixed solution of dimethylsulfoxide and hydroxyethyl starch as a cryoprotectant. In addition, we developed new freezing cases to minimize storage space utilization within the ISS freezer. Finally, we confirmed the recovery of major mononuclear immune cell subsets from the cryopreserved blood cells through a high dimensional analysis of flow cytometric data using 13 cell surface markers. Consequently, this ground study lays the foundation for the cryopreservation of viable blood cells on the ISS, enabling their analysis upon return to Earth. The application of this method in ISS studies will contribute to understanding the impact of space environments on human cells. Moreover, this method may find application in the cryopreservation of blood cells in situations where research facilities are inadequate.