title: High-Throughput Non-Contact Vitrification of Cell-Laden Droplets Based on Cell Printing.
abstract: Cryopreservation is the most promising way for long-term storage of biological samples e.g., single cells and cellular structures. Among various cryopreservation methods, vitrification is advantageous by employing high cooling rate to avoid the formation of harmful ice crystals in cells. Most existing vitrification methods adopt direct contact of cells with liquid nitrogen to obtain high cooling rates, which however causes the potential contamination and difficult cell collection. To address these limitations, we developed a non-contact vitrification device based on an ultra-thin freezing film to achieve high cooling/warming rate and avoid direct contact between cells and liquid nitrogen. A high-throughput cell printer was employed to rapidly generate uniform cell-laden microdroplets into the device, where the microdroplets were hung on one side of the film and then vitrified by pouring the liquid nitrogen onto the other side via boiling heat transfer. Through theoretical and experimental studies on vitrification processes, we demonstrated that our device offers a high cooling/warming rate for vitrification of the NIH 3T3 cells and human adipose-derived stem cells (hASCs) with maintained cell viability and differentiation potential. This non-contact vitrification device provides a novel and effective way to cryopreserve cells at high throughput and avoid the contamination and collection problems.
authors.affiliation: School of Energy and Power Engineering, Xi'an Jiaotong University, Xi'an, 710049, P.R. China.|Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an, 710049, P.R. China.
authors.forename: Meng
authors.initials: M
authors.lastname: Shi
authors.affiliation: Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an, 710049, P.R. China.|State Key Laboratory of Mechanical Structure Strength and Vibration, School of Aerospace, Xi'an Jiaotong University, Xi'an, 710049, P.R. China.
authors.forename: Kai
authors.initials: K
authors.lastname: Ling
authors.affiliation: Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an, 710049, P.R. China.|Department of Biomedical Engineering, Faculty of Engineering, University of Malaya, Lembah Pantai, 50603 Kuala Lumpur, Malaysia.
authors.forename: Kar Wey
authors.initials: KW
authors.lastname: Yong
authors.affiliation: Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an, 710049, P.R. China.|MOE Key Laboratory of Biomedical Information Engineering, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, 710049, P.R. China.
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Li
authors.affiliation: Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an, 710049, P.R. China.|State Key Laboratory of Mechanical Structure Strength and Vibration, School of Aerospace, Xi'an Jiaotong University, Xi'an, 710049, P.R. China.
authors.forename: Shangsheng
authors.initials: S
authors.lastname: Feng
authors.affiliation: Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an, 710049, P.R. China.|MOE Key Laboratory of Biomedical Information Engineering, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, 710049, P.R. China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Biomedical Engineering, Faculty of Engineering, University of Malaya, Lembah Pantai, 50603 Kuala Lumpur, Malaysia.
authors.forename: Belinda
authors.initials: B
authors.lastname: Pingguan-Murphy
authors.affiliation: Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an, 710049, P.R. China.|State Key Laboratory of Mechanical Structure Strength and Vibration, School of Aerospace, Xi'an Jiaotong University, Xi'an, 710049, P.R. China.
authors.forename: Tian Jian
authors.initials: TJ
authors.lastname: Lu
authors.affiliation: Bioinspired Engineering and Biomechanics Center (BEBC), Xi'an Jiaotong University, Xi'an, 710049, P.R. China.|MOE Key Laboratory of Biomedical Information Engineering, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an, 710049, P.R. China.
authors.forename: Feng
authors.initials: F
authors.lastname: Xu
mesh_terms: D000465:Algorithms; D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D002470:Cell Survival; D003198:Computer Simulation; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D051379:Mice; D008962:Models, Theoretical; D041681:NIH 3T3 Cells; D058989:Vitrification