Advanced Biotechnology for Cell Cryopreservation

Abstract

Cell cryopreservation has evolved as an important technology required for supporting various cell-based applications, such as stem cell therapy, tissue engineering, and assisted reproduction. Recent times have witnessed an increase in the clinical demand of these applications, requiring urgent improvements in cell cryopreservation. However, cryopreservation technology suffers from the issues of low cryopreservation efficiency and cryoprotectant (CPA) toxicity. Application of advanced biotechnology tools can significantly improve post-thaw cell survival and reduce or even eliminate the use of organic solvent CPAs, thus promoting the development of cryopreservation. Herein, based on the different cryopreservation mechanisms available, we provide an overview of the applications and achievements of various biotechnology tools used in cell cryopreservation, including trehalose delivery, hydrogel-based cell encapsulation technique, droplet-based cell printing, and nanowarming, and also discuss the associated challenges and perspectives for future development.

Keywords

Cell cryopreservation / Biotechnology / Trehalose delivery / Hydrogel-based cell encapsulation / Droplet-based cell printing / Nanowarming

References

[1] Zhao G, Fu JP. Microfluidics for cryopreservation. Biotechnol Adv, 2017, 35(2): 323-336.

[2] Ballen K. Challenges in umbilical cord blood stem cell banking for stem cell reviews and reports. Stem Cell Rev Rep, 2010, 6(1): 8-14.

[3] Chang A, Kim Y, Hoehn R, et al. Cryopreserved packed red blood cells in surgical patients: past, present, and future. Blood Transfus, 2016, 15(4): 341-347.

[4] Yang J, Pan C, Zhang JM, et al. Exploring the potential of biocompatible osmoprotectants as highly efficient cryoprotectants. ACS Appl Mater Interfaces, 2017, 9(49): 42516-42524.

[5] Gurruchaga H, Saenz del Burgo L, Hernandez RM, et al. Advances in the slow freezing cryopreservation of microencapsulated cells. J Control Release, 2018, 281: 119-138.

[6] Fuller B, Gonzalez-Molina J, Erro E, et al. Applications and optimization of cryopreservation technologies to cellular therapeutics. Cell Gene Therapy Insights, 2017, 3(5): 359-378.

[7] Pal R, Mamidi MK, Das AK, et al. Diverse effects of dimethyl sulfoxide (DMSO) on the differentiation potential of human embryonic stem cells. Arch Toxicol, 2012, 86(4): 651-661.

[8] Shu Z, Heimfeld S, Gao D. Hematopoietic SCT with cryopreserved grafts: adverse reactions after transplantation and cryoprotectant removal before infusion. Bone Marrow Transplant, 2014, 49(4): 469-476.

[9] Elliott GD, Wang SP, Fuller BJ. Cryoprotectants: a review of the actions and applications of cryoprotective solutes that modulate cell recovery from ultra-low temperatures. Cryobiology, 2017, 76: 74-91.

[10] Best BP. Cryoprotectant toxicity: facts, issues, and questions. Rejuvenation Res, 2015, 18(5): 422-436.

[11] Lauterboeck L, Wolkers WF, Glasmacher B. Cryobiological parameters of multipotent stromal cells obtained from different sources. Cryobiology, 2017, 74: 93-102.

[12] Pollock K, Budenske JW, McKenna DH, et al. Algorithm-driven optimization of cryopreservation protocols for transfusion model cell types including Jurkat cells and mesenchymal stem cells. J Tissue Eng Regen Med, 2017, 11(10): 2806-2815.

[13] Mahler S, Desille M, Frémond B, et al. Hypothermic storage and cryopreservation of hepatocytes: the protective effect of alginate gel against cell damages. Cell Transplant, 2003, 12(6): 579-592.

[14] Chinnadurai R, Copland IB, Garcia MA, et al. Cryopreserved mesenchymal stromal cells are susceptible to T-cell mediated apoptosis which is partly rescued by IFNγ licensing. Stem Cells, 2016, 34(9): 2429-2442.

[15] Moll G, Alm JJ, Davies LC, et al. Do cryopreserved mesenchymal stromal cells display impaired immunomodulatory and therapeutic properties?. Stem Cells, 2014, 32(9): 2430-2442.

[16] Shaik S, Wu XY, Gimble J, et al. Effects of decade long freezing storage on adipose derived stem cells functionality. Sci Rep, 2018, 8: 8162.

[17] Zhou QQ, Zhang YL, Zhao M, et al. Mature dendritic cell derived from cryopreserved immature dendritic cell shows impaired homing ability and reduced anti-Viral therapeutic effects. Sci Rep, 2016, 6: 39071.

[18] Sui XJ, Wen CY, Yang J, et al. Betaine combined with membrane stabilizers enables solvent-free whole blood cryopreservation and one-step cryoprotectant removal. ACS Biomater Sci Eng, 2019, 5(2): 1083-1091.

[19] Yang J, Cai NN, Zhai HW, et al. Natural zwitterionic betaine enables cells to survive ultrarapid cryopreservation. Sci Rep, 2016, 6: 37458.

[20] Cao TT, Zhang YQ. The potential of silk sericin protein as a serum substitute or an additive in cell culture and cryopreservation. Amino Acids, 2017, 49(6): 1029-1039.

[21] Kim SM, Yun CK, Park JH, et al. Efficient cryopreservation of human mesenchymal stem cells using silkworm hemolymph-derived proteins. J Tissue Eng Regen Med, 2017, 11(8): 2181-2192.

[22] Tsai S, Chong G, Meng PJ, et al. Sugars as supplemental cryoprotectants for marine organisms. Rev Aquac, 2018, 10: 703-715.

[23] Cottone G. A comparative study of carboxy myoglobin in saccharide–water systems by molecular dynamics simulation. J Phys Chem B, 2007, 111(13): 3563-3569.

[24] He XM, Fowler A, Toner M. Water activity and mobility in solutions of glycerol and small molecular weight sugars: implication for cryo- and lyopreservation. J Appl Phys, 2006, 100(7): 074702.

[25] Sun WQ, Leopold AC, Crowe LM, et al. Stability of dry liposomes in sugar glasses. Biophys J, 1996, 70(4): 1769-1776.

[26] Chen FF, Zhang WJ, Wu W, et al. Cryopreservation of tissue-engineered epithelial sheets in trehalose. Biomaterials, 2011, 32(33): 8426-8435.

[27] Ntai A, La Spada A, de Blasio P, et al. Trehalose to cryopreserve human pluripotent stem cells. Stem Cell Res, 2018, 31: 102-112.

[28] Shinde P, Khan N, Melinkeri S, et al. Freezing of dendritic cells with trehalose as an additive in the conventional freezing medium results in improved recovery after cryopreservation. Transfusion, 2019, 59(2): 686-696.

[29] Stewart S, He XM. Intracellular delivery of trehalose for cell banking. Langmuir, 2019, 35(23): 7414-7422.

[30] Gläfke C, Akhoondi M, Oldenhof H, et al. Cryopreservation of platelets using trehalose: the role of membrane phase behavior during freezing. Biotechnol Progress, 2012, 28(5): 1347-1354.

[31] Zhang M, Oldenhof H, Sieme H, et al. Combining endocytic and freezing-induced trehalose uptake for cryopreservation of mammalian cells. Cryobiology, 2016, 73(3): 412.

[32] Zhang M, Oldenhof H, Sieme H, et al. Freezing-induced uptake of trehalose into mammalian cells facilitates cryopreservation. Biochimica Et Biophys Acta Bba Biomembr, 2016, 1858(6): 1400-1409.

[33] Dovgan B, Barlič A, Knežević M, et al. Cryopreservation of human adipose-derived stem cells in combination with trehalose and reversible electroporation. J Membrane Biol, 2017, 250(1): 1-9.

[34] Dovgan B, Dermol J, Barlič A et al (2016) Cryopreservation of human umbilical stem cells in combination with trehalose and reversible electroporation. 1st World congress on electroporation and pulsed electric fields in biology, medicine and food & environmental technologies. Springer, Singapore, pp 307–310

[35] Shirakashi R, Köstner CM, Müller KJ, et al. Intracellular delivery of trehalose into mammalian cells by electropermeabilization. J Membr Biol, 2002, 189(1): 45-54.

[36] Beattie GM, Crowe JH, Lopez AD, et al. Trehalose: a cryoprotectant that enhances recovery and preserves function of human pancreatic islets after long-term storage. Diabetes, 1997, 46(3): 519-523.

[37] Batista Napotnik T, Miklavčič D. In vitro electroporation detection methods—an overview. Bioelectrochemistry, 2018, 120: 166-182.

[38] Cao YH, Ma EB, Cestellos-Blanco S, et al. Reply to Nathamgari et al. Nanopore electroporation for intracellular delivery of biological macromolecules. Proc Natl Acad Sci USA, 2019, 116(46): 22911.

[39] Rems L, Miklavčič D. Tutorial: electroporation of cells in complex materials and tissue. J Appl Phys, 2016, 119(20): 201101.

[40] Šatkauskas S, Ruzgys P, Venslauskas MS. Towards the mechanisms for efficient gene transfer into cells and tissues by means of cell electroporation. Expert Opin Biol Ther, 2012, 12(3): 275-286.

[41] Yao CG, Liu HM, Zhao YJ, et al. Analysis of dynamic processes in single-cell electroporation and their effects on parameter selection based on the finite-element model. IEEE Trans Plasma Sci, 2017, 45(5): 889-900.

[42] Lynch AL, Chen RJ, Slater NKH. PH-responsive polymers for trehalose loading and desiccation protection of human red blood cells. Biomaterials, 2011, 32(19): 4443-4449.

[43] Mercado SA, Slater NKH. Increased cryosurvival of osteosarcoma cells using an amphipathic pH-responsive polymer for trehalose uptake. Cryobiology, 2016, 73(2): 175-180.

[44] Lynch AL, Chen RJ, Dominowski PJ, et al. Biopolymer mediated trehalose uptake for enhanced erythrocyte cryosurvival. Biomaterials, 2010, 31(23): 6096-6103.

[45] Acker JP, Lu XM, Young V, et al. Measurement of trehalose loading of mammalian cells porated with a metal-actuated switchable pore. Biotechnol Bioeng, 2003, 82(5): 525-532.

[46] Buchanan SS, Gross SA, Acker JP, et al. Cryopreservation of stem cells using trehalose: evaluation of the method using a human hematopoietic cell line. Stem Cells Dev, 2004, 13(3): 295-305.

[47] Eroglu A, Russo MJ, Bieganski R, et al. Intracellular trehalose improves the survival of cryopreserved mammalian cells. Nat Biotechnol, 2000, 18(2): 163-167.

[48] Rao W, Huang HS, Wang H, et al. Nanoparticle-mediated intracellular delivery enables cryopreservation of human adipose-derived stem cells using trehalose as the sole cryoprotectant. ACS Appl Mater Interfaces, 2015, 7(8): 5017-5028.

[49] Stefanic M, Ward K, Tawfik H, et al. Apatite nanoparticles strongly improve red blood cell cryopreservation by mediating trehalose delivery via enhanced membrane permeation. Biomaterials, 2017, 140: 138-149.

[50] Zhang WJ, Rong JH, Wang Q, et al. The encapsulation and intracellular delivery of trehalose using a thermally responsive nanocapsule. Nanotechnology, 2009, 20(27): 275101.

[51] Russo MJ, Bayley H, Toner M. Reversible permeabilization of plasma membranes with an engineered switchable pore. Nat Biotechnol, 1997, 15(3): 278-282.

[52] Solanki A, Kim JD, Lee KB. Nanotechnology for regenerative medicine: nanomaterials for stem cell imaging. Nanomedicine, 2008, 3(4): 567-578.

[53] Vernekar VN, James R, Smith KJ, et al. Nanotechnology applications in stem cell science for regenerative engineering. J Nanosci Nanotechnol, 2016, 16(9): 8953-8965.

[54] Watermann A, Brieger J. Mesoporous silica nanoparticles as drug delivery vehicles in cancer. Nanomaterials, 2017, 7(7): 189.

[55] Qian HQ, Liu BR, Jiang XQ. Application of nanomaterials in cancer immunotherapy. Mater Today Chem, 2018, 7: 53-64.

[56] Orive G, Hernández RM, Gascón AR, et al. Cell encapsulation: promise and progress. Nat Med, 2003, 9(1): 104-107.

[57] Zimmermann H, Ehrhart F, Zimmermann D, et al. Hydrogel-based encapsulation of biological, functional tissue: fundamentals, technologies and applications. Appl Phys A, 2007, 89(4): 909-922.

[58] Kang A, Park J, Ju J, et al. Cell encapsulation via microtechnologies. Biomaterials, 2014, 35(9): 2651-2663.

[59] Zhang W, He X. Microencapsulating and banking living cells for cell-based medicine. J Healthc Eng, 2011, 2(4): 427-446.

[60] Huang HS, He XM. Microscale materials and devices for cell cryopreservation by vitrification, 2016, Singapore: Multiscale Technologies for Cryomedicine, World Scientific, 101-124.

[61] Dluska E, Cui ZF, Markowska-Radomska A, et al. Cryoprotection and banking of living cells in a 3D multiple emulsion-based carrier. Biotechnol J, 2017, 12(8): 1600692.

[62] Majewski RL, Zhang WJ, Ma XJ, et al. Bioencapsulation technologies in tissue engineering. J Appl Biomater Funct Mater, 2016, 14(4): e395-e403.

[63] Rabanel JM, Banquy X, Zouaoui H, et al. Progress technology in microencapsulation methods for cell therapy. Biotechnol Prog, 2009, 25(4): 946-963.

[64] Chopra P, Nayak D, Nanda A, et al. Fabrication of poly(vinyl alcohol)-carrageenan scaffolds for cryopreservation: effect of composition on cell viability. Carbohydr Polym, 2016, 147: 509-516.

[65] Vrana NE, Matsumura K, Hyon SH, et al. Cell encapsulation and cryostorage in PVA–gelatin cryogels: incorporation of carboxylated ε-poly-l-lysine as cryoprotectant. J Tissue Eng Regen Med, 2011, 6(4): 280-290.

[66] Vrana NE, O’Grady A, Kay E, et al. Cell encapsulation within PVA-based hydrogels via freeze-thawing: a one-step scaffold formation and cell storage technique. J Tissue Eng Regen Med, 2009, 3(7): 567-572.

[67] Zeng J, Yin YX, Zhang L, et al. A supramolecular gel approach to minimize the neural cell damage during cryopreservation process. Macromol Biosci, 2016, 16(3): 363-370.

[68] Jain M, Rajan R, Hyon SH, et al. Hydrogelation of dextran-based polyampholytes with cryoprotective properties via click chemistry. Biomater Sci, 2014, 2(3): 308-317.

[69] Popa EG, Rodrigues MT, Coutinho DF, et al. Cryopreservation of cell laden natural origin hydrogels for cartilage regeneration strategies. Soft Matter, 2013, 9(3): 875-885.

[70] Wolters GHJ, Fritschy WM, Gerrits D, et al. A versatile alginate droplet generator applicable for microencapsulation of pancreatic islets. J Appl Biomater, 1992, 3(4): 281-286.

[71] Malpique R, Osório LM, Ferreira DS, et al. Alginate encapsulation as a novel strategy for the cryopreservation of neurospheres. Tissue Eng Part C Methods, 2010, 16(5): 965-977.

[72] Zhang WJ, He XM. Encapsulation of living cells in small (~ 100 μm) alginate microcapsules by electrostatic spraying: a parametric study. J Biomech Eng, 2009, 131(7): 074515.

[73] Zhang WJ, Yang GE, Zhang AL, et al. Preferential vitrification of water in small alginate microcapsules significantly augments cell cryopreservation by vitrification. Biomed Microdevices, 2010, 12(1): 89-96.

[74] Zhao ST, Agarwal P, Rao W, et al. Coaxial electrospray of liquid core–hydrogel shell microcapsules for encapsulation and miniaturized 3D culture of pluripotent stem cells. Integr Biol, 2014, 6(9): 874-884.

[75] Lu YC, Fu DJ, An D, et al. Scalable production and cryostorage of organoids using core-shell decoupled hydrogel capsules. Adv Biosyst, 2017, 1(12): 1700165.

[76] Naqvi SM, Gansau J, Buckley CT. Priming and cryopreservation of microencapsulated marrow stromal cells as a strategy for intervertebral disc regeneration. Biomed Mater, 2018, 13(3): 034106.

[77] Kashaninejad N, Shiddiky MJA, Nguyen NT. Advances in microfluidics-based assisted reproductive technology: from sperm sorter to reproductive system-on-a-chip. Adv Biosyst, 2018, 2(3): 1700197.

[78] Chen WY, Shu ZQ, Gao DY, et al. Sensing and sensibility: single-islet-based quality control assay of cryopreserved pancreatic islets with functionalized hydrogel microcapsules. Adv Healthcare Mater, 2016, 5(2): 223-231.

[79] Huang HS, Choi JK, Rao W, et al. Alginate hydrogel microencapsulation inhibits devitrification and enables large-volume low-CPA cell vitrification. Adv Funct Mater, 2015, 25(44): 6839-6850.

[80] Zhao G, Liu XL, Zhu KX, et al. Hydrogel encapsulation facilitates rapid-cooling cryopreservation of stem cell-laden core-shell microcapsules as cell-biomaterial constructs. Adv Healthcare Mater, 2017, 6(23): 1700988.

[81] Nir R, Lamed R, Gueta L, et al. Single-cell entrapment and microcolony development within uniform microspheres amenable to flow cytometry. Appl Environ Microbiol, 1990, 56: 2870-2875.

[82] Perçin G, Yaralioglu GG, Khuri-Yakub BT. Micromachined droplet ejector arrays. Rev Sci Instrum, 2002, 73(12): 4385-4389.

[83] Blossey R. Self-cleaning surfaces: virtual realities. Nat Mater, 2003, 2(5): 301-306.

[84] Demirci U. Acoustic picoliter droplets for emerging applications in semiconductor industry and biotechnology. J Microelectromech Syst, 2006, 15(4): 957-966.

[85] Reis N, Ainsley C, Derby B. Ink-jet delivery of particle suspensions by piezoelectric droplet ejectors. J Appl Phys, 2005, 97(9): 094903.

[86] de Vries RJ, Banik PD, Nagpal S, et al. Bulk droplet vitrification: an approach to improve large-scale hepatocyte cryopreservation outcome. Langmuir, 2019, 35(23): 7354-7363.

[87] Youn JR, Song YS. Cell-encapsulating droplet formation and freezing. Appl Phys Lett, 2012, 101(13): 133701.

[88] Demirci U, Montesano G. Cell encapsulating droplet vitrification. Lab Chip, 2007, 7(11): 1428.

[89] El Assal R, Guven S, Gurkan UA, et al. Bio-inspired cryo-ink preserves red blood cell phenotype and function during nanoliter vitrification. Adv Mater, 2014, 26(33): 5815-5822.

[90] Shi M, Ling K, Yong KW, et al. High-throughput non-contact vitrification of cell-laden droplets based on cell printing. Sci Rep, 2015, 5: 17928.

[91] Akiyama Y, Shinose M, Watanabe H, et al. Cryoprotectant-free cryopreservation of mammalian cells by superflash freezing. Proc Natl Acad Sci USA, 2019, 116(16): 7738-7743.

[92] Deller RC, Pessin JE, Vatish M, et al. Enhanced non-vitreous cryopreservation of immortalized and primary cells by ice-growth inhibiting polymers. Biomater Sci, 2016, 4(7): 1079-1084.

[93] Manuchehrabadi N, Gao Z, Zhang JJ, et al. Improved tissue cryopreservation using inductive heating of magnetic nanoparticles. Sci Transl Med, 2017, 9(379): eaah4586.

[94] Czajka C. Nanowarming improves cryopreservation. Science, 2017, 355(6328): 920-921.

[95] Etheridge ML, Xu Y, Rott L, et al. RF heating of magnetic nanoparticles improves the thawing of cryopreserved biomaterials. Technology, 2014, 2(3): 229-242.

[96] Wang T, Zhao G, Deng ZS, et al. Theoretical investigation of a novel microwave antenna aided cryovial for rapid and uniform rewarming of frozen cryoprotective agent solutions. Appl Therm Eng, 2015, 89: 968-977.

[97] Liu XL, Zhao G, Chen ZR, et al. Dual suppression effect of magnetic induction heating and microencapsulation on ice crystallization enables low-cryoprotectant vitrification of stem cell–Alginate hydrogel constructs. ACS Appl Mater Interfaces, 2018, 10(19): 16822-16835.

[98] Pan JJ, Shu ZQ, Zhao G, et al. Towards uniform and fast rewarming for cryopreservation with electromagnetic resonance cavity: numerical simulation and experimental investigation. Appl Therm Eng, 2018, 140: 787-798.

[99] Khosla K, Wang YR, Hagedorn M, et al. Gold nanorod induced warming of embryos from the cryogenic state enhances viability. ACS Nano, 2017, 11(8): 7869-7878.

[100] Wang JY, Zhao G, Zhang ZL, et al. Magnetic induction heating of superparamagnetic nanoparticles during rewarming augments the recovery of hUCM-MSCs cryopreserved by vitrification. Acta Biomater, 2016, 33: 264-274.

[101] Khosla K, Zhan L, Bhati A, et al. Characterization of laser gold nanowarming: a platform for millimeter-scale cryopreservation. Langmuir, 2019, 35(23): 7364-7375.

[102] Panhwar F, Chen ZR, Hossain SMC, et al. Near-infrared laser mediated modulation of ice crystallization by two-dimensional nanosheets enables high-survival recovery of biological cells from cryogenic temperatures. Nanoscale, 2018, 10(25): 11760-11774.

[103] Swioklo S, Constantinescu A, Connon CJ. Alginate-encapsulation for the improved hypothermic preservation of human adipose-derived stem cells. Stem Cells Transl Med, 2016, 5(3): 339-349.

[104] Swioklo S, Ding P, Pacek AW, et al. Process parameters for the high-scale production of alginate-encapsulated stem cells for storage and distribution throughout the cell therapy supply chain. Process Biochem, 2017, 59: 289-296.

[105] Bissoyi A, Nayak B, Pramanik K, et al. Targeting cryopreservation-induced cell death: a review. Biopreservation Biobanking, 2014, 12(1): 23-34.

[106] Baust JG, Snyder KK, van Buskirk R, et al. Integrating molecular control to improve cryopreservation outcome. Biopreservation Biobanking, 2017, 15(2): 134-141.