DOCSLIB.ORG

DNA Origami As Emerging Technology for the Engineering of Fluorescent and Plasmonic-Based Biosensors

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
DNA Origami As Emerging Technology for the Engineering of Fluorescent and Plasmonic-Based Biosensors materials Review DNA Origami as Emerging Technology for the Engineering of Fluorescent and Plasmonic-Based Biosensors 1, , 2, , 1 2, Morgane Loretan * y, Ivana Domljanovic * y, Mathias Lakatos , Curzio Rüegg y and 1, Guillermo P. Acuna y 1 Photonic Nanosystems, Department of Physics, Faculty of Science and Medicine, University of Fribourg, Chemin du Musée 3, PER08, 1700 Fribourg, Switzerland; [email protected] (M.L.); [email protected] (G.P.A.) 2 Laboratory of Experimental and Translational Oncology, Department of Oncology, Microbiology and Immunology, Faculty of Science and Medicine, University of Fribourg, Chemin du Musée 18, PER17, 1700 Fribourg, Switzerland; [email protected] * Correspondence: [email protected] (M.L.); [email protected] (I.D.) These authors contributed equally to this work. y Received: 7 April 2020; Accepted: 5 May 2020; Published: 9 May 2020 Abstract: DNA nanotechnology is a powerful and promising tool for the development of nanoscale devices for numerous and diverse applications. One of the greatest potential fields of application for DNA nanotechnology is in biomedicine, in particular biosensing. Thanks to the control over their size, shape, and fabrication, DNA origami represents a unique opportunity to assemble dynamic and complex devices with precise and predictable structural characteristics. Combined with the addressability and flexibility of the chemistry for DNA functionalization, DNA origami allows the precise design of sensors capable of detecting a large range of different targets, encompassing RNA, DNA, proteins, small molecules, or changes in physico-chemical parameters, that could serve as diagnostic tools. Here, we review some recent, salient developments in DNA origami-based sensors centered on optical detection methods (readout) with a special emphasis on the sensitivity, the selectivity, and response time. We also discuss challenges that still need to be addressed before this approach can be translated into robust diagnostic devices for bio-medical applications. Keywords: DNA nanotechnology; DNA origami; biosensors; optics (plasmonic and fluorescence sensing) 1. Introduction 1.1. DNA Nanotechnology Nanotechnology allows applications of novel materials and devices at the nanoscale level in all disciplines of science. One emerging material for nanoscale engineering is DNA [1]. Some of the main characteristics of DNA are the high specificity, programmability, flexibility, and sub-nanometer precision. These characteristics allow DNA to be used for the construction of highly precise nanostructures, as demonstrated first by Seeman, in his ground-breaking research in the early 1980s [2]. Self-assembly of DNA nanostructures started with crossovers and branched junctions [3] followed by a variety of DNA objects such as polyhedral [4], prisms [5], and buckyballs [4]. Subsequently, self-assembly of DNA was further developed into an advanced order of design using DNA crossover tiles [6]. With this strategy, sticky ends are added to DNA for creating two-dimensional (2D) [7] and three-dimensional (3D) [8,9] periodic lattices. Another more advanced single-stranded DNA tiles strategy for DNA-based construction is DNA bricks that form large DNA structures in 2D and 3D [10]. All self-assemblies Materials 2020, 13, 2185; doi:10.3390/ma13092185 www.mdpi.com/journal/materials Materials 2020, 13, x FOR PEER REVIEW 2 of 25 Materials 2020, 13, 2185 2 of 24 structures in 2D and 3D [10]. All self-assemblies mentioned above are based on a combination of “short”mentioned single above-stranded are based DNA on aor combination pre-assembled of “short” DNA single-strandedmotifs. The structures DNA or assembled pre-assembled via these DNA methodsmotifs. The can structures be used as assembled programmable via these scaffo methodslds forcan the beorganization used as programmable of different nanoparticles scaffolds forthe or moleculesorganization [11] of, but diff areerent still nanoparticles limited by the or size molecules and the [11 complexity.], but are still In 2006, limited a method by the sizecalled and DNA the origamicomplexity. was Inintroduced 2006, a method by Rothemund called DNA [12] origami. This method was introduced enabled an by increase Rothemund in complexity [12]. This methodand the creatienabledon anof increaselarger nanostructures in complexity and(in the the range creation of hundreds of larger nanostructures of nm) [13,14] (in. The the technique range of hundreds consists of anm) long [13 single,14].- Thestranded technique DNA consists scaffold of (usually a long single-stranded from the genome DNA of the sca bacteriophageffold (usually fromM13, the containing genome approximatelyof the bacteriophage seven M13,to eight containing thousand approximately nucleotides), sevenwhich to is eight folded thousand into desired nucleotides), shapes with which the is helpfolded of intoa set desiredof hundreds shapes of withshort the complementary help of a set ofstaple hundreds strands of (approximately short complementary 40 nucleotides staple strands long). One(approximately of the advantages 40 nucleotides of the long).DNA Oneorigami of the method advantages compared of the to DNA other origami assembly method strategies compared is the to simplicityother assembly of using strategies a scaffold is the strand simplicity and offolding using it a scaintoff oldany strand pre-designed and folding shape, it into and any that pre-designed it does not needshape, a andstoichiometric that it does blend not need of component a stoichiometric strands blend [15] of. The component scaffold strandshas a well [15-].known The sca nucleotidesffold has a well-knownsequence, whereas nucleotides complementary sequence, whereasnucleobases complementary of the staples nucleobases are carefully of chosen the staples depending are carefully on the wantedchosen dependingdesign of the on theDNA wanted origami design structure. of the Upon DNA origamihybridization structure. of the Upon scaffold hybridization with the staple of the strands,scaffold withevery the nucleotide staple strands, of each every DNA nucleotide staple will of then each “take DNA the staple right will place” then by “take hybridizing the rightplace” with the by complementaryhybridizing with sequence the complementary on the scaffold. sequence Ther onefore, the scatheff “folding”old. Therefore, of the the scaffold “folding” into ofthe the designed scaffold specificinto the shape designed takes specific place in shape solution takes under place temperature in solution and under salt temperature concentration and conditions salt concentration ensuring theconditions complete ensuring annealing the completeof all strands annealing (Figure of 1). all This strands technique (Figure 1allows). This the technique design allowsof highly the diverse, design complex,of highly diverse,multi-functional, complex, 2D, multi-functional, and 3D structures 2D, and[16– 3D24]. structures [16–24]. Figure 1. Principle of the DNA origami assembly. The DNA origami method consists of a long Figuresingle-strand 1. Principle DNA of (“sca the ffDNAold”) origami and several assembly. hundreds The DNA of short origami ssDNA method strands consists (“staples”). of a long During single an- strandannealing DNA process, (“scaffold”) the “staples” and several fold the hundreds “scaffold” of into short 2D orssDNA 3D structures. strands (“staples”). During an annealing process, the “staples” fold the “scaffold” into 2D or 3D structures. The fact that the whole sequence is known, together with the ability to attach functional molecules to singleThe strandsfact that in DNAthe whole origami sequence structures is withknown, a high together precision with and the accuracy, ability makes to attach DNA functional origami a moleculeshighly addressable to single tool strands with predictablein DNA origami conformation structures structures with a [high25]. Besides,precision DNA and origami accuracy, structures makes DNAare natural origami biopolymers a highly addressable that are biocompatible, tool with predictable biodegradable, conformation and minimally structures toxic [25]. [ 26Besides]. Thus,, DNA this origamiapproach structures has a great are potential natural biopolymers in biomedical tha applicationst are biocompatible, [26,27]. Furthermore, biodegradable, it was and shown minimally that toxicDNA [2 origami6]. Thus, structures this approach are more has a stable great than potential other in nucleic biomedical acid structures, applications facilitating [26,27]. Furthermore, their use for itbiological was shown and technicalthat DNA applications origami structures [28]. In addition, are more the stable development than other of DNA nucleic origami acid enabledstructures, the facilitatielaborationng their of “dynamic” use for biological structures and whose technical functions applications rely upon [28 conformational]. In addition, the changes development induced byof DNAexternal origami stimuli, enabled externally the applied elaboration fields, orof strand-displacement “dynamic” structures reactions whose [29 ].functions The conformational rely upon conformationalchanges may be changes then exploited induced to by generate external measurable stimuli, externally signals toapplied “sense” fields the, originalor strand stimulus-displacement [29,30]. reactionsThe dynamic [29].structures The conformational can be used changes in diff erentmay be applications then exploited [31– 34to], generate in particular measurable for biomolecular signals to “sense”sensing. the For
Load more

Recommended publications
  • CRYOPRESERVATION 2021 Research and Resources ORIP’S MISSION ORIP Advances the NIH Mission by Supporting Orip.Nih.Gov Infrastructure for Innovation
    OFFICE OF RESEARCH INFRASTRUCTURE PROGRAMS CRYOPRESERVATION 2021 Research and Resources ORIP’S MISSION ORIP advances the NIH mission by supporting orip.nih.gov infrastructure for innovation. This support is focused on research resources, including animal twitter.com/NIH_ORIP models for human diseases, cutting-edge scientific instrumentation, construction and modernization of research facilities, and research training Program Contacts: opportunities for veterinary scientists. Through Miguel Contreras, Ph.D. Desiree von Kollmar continued engagement with NIH Institutes, [email protected] [email protected] Centers, and Offices and the biomedical research Phone: 301-435-0045 Phone: 301-594-9410 community, ORIP empowers and expands existing Oleg Mirochnitchenko, Ph.D. Sige Zou, Ph.D. programs and develops new initiatives to support [email protected] [email protected] NIH research at the forefront of scientific progress. Phone: 301-435-0748 Phone: 301-435-0749 projects on developing efficient OVERVIEW germplasm preservation methods, including cryopreservation, for diverse Animal models are essential to understanding diseases and animal models. These projects maintaining health of humans through development of are improving existing methods diagnostic approaches and therapeutic interventions. These to preserve embryos (including disease models are being generated at an unprecedented NHP embryos), eggs, or sperm rate due to rapidly evolving technological advancements, for a variety of animal model such as gene editing. This rapid increase in animal models, species or are developing new however, is creating challenges in how to maintain these preservation methods, such critical resources in reliable and cost-effective ways. Long- as for zebrafish or Drosophila term preservation of the genetic stock of such models is melanogaster embryos.
    [Show full text]
  • Molecular Beacon Nanosensors for Live Cell Detection and Tracking Differentiation and Reprogramming
    Downloaded from orbit.dtu.dk on: Oct 03, 2021 Molecular beacon nanosensors for live cell detection and tracking differentiation and reprogramming Ilieva, Mirolyuba Publication date: 2013 Document Version Publisher's PDF, also known as Version of record Link back to DTU Orbit Citation (APA): Ilieva, M. (2013). Molecular beacon nanosensors for live cell detection and tracking differentiation and reprogramming. General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. Users may download and print one copy of any publication from the public portal for the purpose of private study or research. You may not further distribute the material or use it for any profit-making activity or commercial gain You may freely distribute the URL identifying the publication in the public portal If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Molecular beacon nanosensors for live cell detection and tracking differentiation and reprogramming Mirolyuba Ilieva PhD Thesis April 2013 Mirolyuba Simeonova Ilieva Molecular beacon nanosensors for live cell detection and tracking differentiation and reprogramming PhD Thesis April 2013 Supervisor Assoc. Prof. Martin Dufva Co-supervisor Professor Jenny Emnéus Department of Micro- and Nanotechnology Technical University of Denmark Abstract High-sensitive and high-affinity methods to measure gene expression inside living cells have proven to be invaluable in regards to understanding fundamental processes such as cell differentiation, reprogramming, regeneration and cancer genesis.
    [Show full text]
  • Programming Structured DNA Assemblies to Probe Biophysical Processes
    Programming Structured DNA Assemblies to Probe Biophysical Processes The MIT Faculty has made this article openly available. Please share how this access benefits you. Your story matters. Citation Wamhoff, Eike-Christian et al. “Programming Structured DNA Assemblies to Probe Biophysical Processes.” Annual Review of Biophysics 48 (2019): 395-419 © 2019 The Author(s) As Published 10.1146/ANNUREV-BIOPHYS-052118-115259 Publisher Annual Reviews Version Author's final manuscript Citable link https://hdl.handle.net/1721.1/125280 Terms of Use Creative Commons Attribution-Noncommercial-Share Alike Detailed Terms http://creativecommons.org/licenses/by-nc-sa/4.0/ HHS Public Access Author manuscript Author ManuscriptAuthor Manuscript Author Annu Rev Manuscript Author Biophys. Author Manuscript Author manuscript; available in PMC 2020 February 22. Published in final edited form as: Annu Rev Biophys. 2019 May 06; 48: 395–419. doi:10.1146/annurev-biophys-052118-115259. Programming Structured DNA Assemblies to Probe Biophysical Processes Eike-Christian Wamhoff, James L. Banal, William P. Bricker, Tyson R. Shepherd, Molly F. Parsons, Rémi Veneziano, Matthew B. Stone, Hyungmin Jun, Xiao Wang, Mark Bathe Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA; Abstract Structural DNA nanotechnology is beginning to emerge as a widely accessible research tool to mechanistically study diverse biophysical processes. Enabled by scaffolded DNA origami in which a long single strand of DNA is weaved throughout an entire target nucleic acid assembly to ensure its proper folding, assemblies of nearly any geometric shape can now be programmed in a fully automatic manner to interface with biology on the 1–100-nm scale.
    [Show full text]
  • Bottom-Up Self-Assembly Based on DNA Nanotechnology
    nanomaterials Review Bottom-Up Self-Assembly Based on DNA Nanotechnology 1, 1, 1 1 1,2,3, Xuehui Yan y, Shujing Huang y, Yong Wang , Yuanyuan Tang and Ye Tian * 1 College of Engineering and Applied Sciences, State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University, Nanjing 210023, China; [email protected] (X.Y.); [email protected] (S.H.); [email protected] (Y.W.); [email protected] (Y.T.) 2 Shenzhen Research Institute of Nanjing University, Shenzhen 518000, China 3 Chemistry and Biomedicine Innovation Center, Nanjing University, Nanjing 210023, China * Correspondence: [email protected] These authors contributed equally to this work. y Received: 9 September 2020; Accepted: 12 October 2020; Published: 16 October 2020 Abstract: Manipulating materials at the atomic scale is one of the goals of the development of chemistry and materials science, as it provides the possibility to customize material properties; however, it still remains a huge challenge. Using DNA self-assembly, materials can be controlled at the nano scale to achieve atomic- or nano-scaled fabrication. The programmability and addressability of DNA molecules can be applied to realize the self-assembly of materials from the bottom-up, which is called DNA nanotechnology. DNA nanotechnology does not focus on the biological functions of DNA molecules, but combines them into motifs, and then assembles these motifs to form ordered two-dimensional (2D) or three-dimensional (3D) lattices. These lattices can serve as general templates to regulate the assembly of guest materials. In this review, we introduce three typical DNA self-assembly strategies in this field and highlight the significant progress of each.
    [Show full text]
  • Cryopreservation of Mouse Resources Toru Takeo1* , Satohiro Nakao1, Yoshiko Nakagawa1, Jorge M
    Takeo et al. Laboratory Animal Research (2020) 36:33 Laboratory Animal Research https://doi.org/10.1186/s42826-020-00066-w REVIEW Open Access Cryopreservation of mouse resources Toru Takeo1* , Satohiro Nakao1, Yoshiko Nakagawa1, Jorge M. Sztein1 and Naomi Nakagata2 Abstract The cryopreservation of sperm and embryos is useful to efficiently archive valuable resources of genetically engineered mice. Till date, more than 60,000 strains of genetically engineered mice have been archived in mouse banks worldwide. Researchers can request for the archived mouse strains for their research projects. The research infrastructure of mouse banks improves the availability of mouse resources, the productivity of research projects, and the reproducibility of animal experiments. Our research team manages the mouse bank at the Center for Animal Resources and Development in Kumamoto University and continuously develops new techniques in mouse reproductive technology to efficiently improve the system of mouse banking. In this review, we introduce the activities of mouse banks and the latest techniques used in mouse reproductive technology. Keywords: Genetically engineered mice, Mouse bank, Reproductive technology, Sperm, Embryo, Cryopreservation, In vitro fertilization, Cold storage, Hands-on workshop Introduction Resource Database (CARD R-BASE, http://cardb.cc.ku- A genetically engineered mouse is a powerful tool to eluci- mamoto-u.ac.jp/transgenic/). Till date, the international date the complex communications between genes or organs collaboration of mouse banks (International Mouse in health and diseases [1]. Moreover, humanized mouse Strain Resource: IMSR) has successfully collected more models derived from immunosuppressed mice are helpful than 60,000 strains of genetically engineered mice to bridge the gap of discovery and the development of new (Table 1).
    [Show full text]
  • Using Modular Preformed DNA Origami Building Blocks to Fold Dynamic 3D Structures
    Using Modular Preformed DNA Origami Building Blocks to Fold Dynamic 3D Structures Thesis Presented in Partial Fulfillment of the Requirements for the Degree Master of Science in the Graduate School of The Ohio State University By Gunter Eickert, B.S. Graduate Program in Mechanical Engineering The Ohio State University 2014 Thesis Committee: Carlos Castro, Advisor Haijun Su Copyright by Gunter Erick Eickert 2014 Abstract DNA origami is a bottom-up approach that takes advantage of DNA’s structure and lock key sequencing to build nano scale machines and structures. By introducing specific single stranded DNA (ssDNA) “staples”, a ssDNA “scaffold” can be folded into a desired 2D or 3D structure. Using this method, a variety of shapes have been formed, including tetrahedrons and octahedrons. These structures have been explored as drug carriers, enzyme platforms, signal markers, and for other medical and research uses. However, each of these structures must be specifically designed and often pertain to only one particular application. A DNA origami structure that functions as a building block was designed to allow for the creation of several different 3D structures without the need for a lengthy design process. An annealing process called a thermal ramp was then used to fold the structures. The building block was made of four equilateral triangles arranged into a parallelogram. This is ideal since a parallelogram can be used to form three of the five platonic solids, in addition to many non-platonic shapes. The parallelogram was successfully folded into a tetrahedron and an octahedron by introducing staples that bound to overhangs on its edges.
    [Show full text]
  • Infertility Services
    Medical Policy Assisted Reproductive Services/Infertility Services Document Number: 002 *Commercial and Qualified Health Plans MassHealth Authorization required X No notification or authorization Not covered X *Not all commercial plans cover this service, please check plan’s benefit package to verify coverage. Contents Overview ....................................................................................................................................................... 2 Coverage Guidelines ..................................................................................................................................... 2 MassHealth, and Certain Custom Plans ........................................................................................................ 2 Covered Services/Procedures ....................................................................................................................... 3 General Eligibility Coverage Criteria ............................................................................................................. 3 SERVICE -SPECIFIC INFERTILITY COVERAGE FOR MEMBERS WITH UTERI and OVARIES ............................... 5 Artificial Insemination (AI)/Intrauterine Insemination (IUI) ......................................................................... 5 Conversion from IUI to In Vitro Fertilization (IVF) ........................................................................................ 6 In Vitro Fertilization (IVF) for Infertility .......................................................................................................
    [Show full text]
  • CRYOPRESERVATION Cryoprotectant Chemicals an Integral Part of Cell Culture Involves the Successful Preservation of Im- Portant Cell Lines
    CRYOPRESERVATION Cryoprotectant Chemicals An integral part of cell culture involves the successful preservation of im- portant cell lines. A primary factor for success is developing an appropri- DIMETHYL SULFOXIDE 25 ml 8.90 196055 100 ml 15.40 ate protocol for a particular cell line. A common misconception among RT [67-68-5] (DMSO) 500 ml 27.40 scientists is that preserving methods for a given cell type are transfer- Cell Culture Reagent 1 liter 45.60 able to similar cell types. Although, this may be true on occassion, it is Purity: ≥99% far from the general rule. Protocols require modification for each cell Ideal for use as a cryoprotectant. NOT FOR HUMAN USE! type. Another key factor for successful preservation is the use of an ef- C2H6SO MW 78.13 fective cryoprotectant. The discovery of glycerol as a preservation me- ETHYLENE GLYCOL 250 ml 10.80 dium led to the rapid discovery of other protectants such as methanol, 151089 1 liter 22.50 RT [107-21-1] ethylene glycol, and DMSO (dimethyl sulfoxide). Purity: 99% min. 1 ml = approx. 1.11 g With any cryopreservation process, the primary objective is maximum C2H6O2 MW 62.1 recovery with minimal damage to the cells. The absence of an effective D-(+)-GLUCOSE 100 g 15.00 cryoprotectant leads to the formation of intracellular ice crystals which 194672 1 kg 23.50 RT [50-99-7] puncture organelles and cell membranes. Also, as water within the me- (Dextrose; Corn sugar) 5 kg 94.00 dium freezes, cells become dehydrated altering the salt concentration Cell Culture Reagent Anhydrous and causing morphological or biological changes after recovery.
    [Show full text]
  • Effects of Sample Treatments on Genome Recovery Via Single-Cell Genomics
    The ISME Journal (2014) 8, 2546–2549 & 2014 International Society for Microbial Ecology All rights reserved 1751-7362/14 www.nature.com/ismej SHORT COMMUNICATION Effects of sample treatments on genome recovery via single-cell genomics Scott Clingenpeel1, Patrick Schwientek1, Philip Hugenholtz2 and Tanja Woyke1 1DOE Joint Genome Institute, Walnut Creek, CA, USA and 2Australian Centre for Ecogenomics, School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, Queensland, Australia Single-cell genomics is a powerful tool for accessing genetic information from uncultivated microorganisms. Methods of handling samples before single-cell genomic amplification may affect the quality of the genomes obtained. Using three bacterial strains we show that, compared to cryopreservation, lower-quality single-cell genomes are recovered when the sample is preserved in ethanol or if the sample undergoes fluorescence in situ hybridization, while sample preservation in paraformaldehyde renders it completely unsuitable for sequencing. The ISME Journal (2014) 8, 2546–2549; doi:10.1038/ismej.2014.92; published online 13 June 2014 Relatively little is known about the functioning of genomes from single cells. Three methods of sample complex microbial communities, largely due to the preservation were tested: cryopreservation with difficulty in culturing most microbes (Rappe and 20% glycerol as a cryoprotectant, preservation in Giovannoni, 2003). Although metagenomics can 70% ethanol, and preservation in 4% paraformalde- provide information on the genetic capabilities of hyde. Because paraformaldehyde treatment causes the entire community, it is difficult to connect crosslinks between nucleic acids and proteins, we predicted gene functions to specific organisms using tested cells exposed to 4% paraformaldehyde with metagenomics (Morales and Holben, 2011).
    [Show full text]
  • Use of Machine Learning with Temporal Photoluminescence Signals from Cdte Quantum Dots for Temperature Measurement in Microfluidic Devices Charles Lewis, James W
    www.acsanm.org Article Use of Machine Learning with Temporal Photoluminescence Signals from CdTe Quantum Dots for Temperature Measurement in Microfluidic Devices Charles Lewis, James W. Erikson, Derek A. Sanchez, C. Emma McClure, Gregory P. Nordin, Troy R. Munro, and John S. Colton* Cite This: ACS Appl. Nano Mater. 2020, 3, 4045−4053 Read Online ACCESS Metrics & More Article Recommendations ABSTRACT: Because of the vital role of temperature in many biological processes studied in microfluidic devices, there is a need to develop improved temperature sensors and data analysis algorithms. The photoluminescence (PL) of nanocrystals (quan- tum dots) has been successfully used in microfluidic temperature devices, but the accuracy of the reconstructed temperature has been limited to about 1 K over a temperature range of tens of degrees. A machine learning algorithm consisting of a fully connected network of seven layers with decreasing numbers of nodes was developed and applied to a combination of normalized spectral and time-resolved PL data of CdTe quantum dot emission in a microfluidic device. The data used by the algorithm were collected over two temperature ranges: 10−300 K and 298−319 K. The accuracy of each neural network was assessed via a mean absolute error of a holdout set of data. For the low-temperature regime, the accuracy was 7.7 K, or 0.4 K when the holdout set is restricted to temperatures above 100 K. For the high-temperature regime, the accuracy was 0.1 K. This method provides demonstrates a potential machine learning approach to accurately sense temperature in microfluidic (and potentially nanofluidic) devices when the data analysis is based on normalized PL data when it is stable over time.
    [Show full text]
  • Cryopreservation, It Will Chill You. Laura Pérez, Adolfo Rodríguez, Carlos Saúco
    Cryopreservation, it will chill you. Laura Pérez, Adolfo Rodríguez, Carlos Saúco Abstract— Tardigrades are micro-animals capable of undergoing cryptobiosis, a freezing process that allows them to survive in damaging environments. Scientists have taken advantage of this capacity for cryopreservation, conserving cells and tissues in time using different techniques such as vitrification for embryos, encapsulation-dehydration for plants or profound hypothermia in surgery. The mechanism is ruled by thermodynamics processes that follow a loss of water by a chemical potential gradient. Key words— Cryopreservation, Cryptobiosis, Encapsulation-Dehydration, Freezing, Tardigrade, Recrystallisation, Vitrification —————————— ◆ —————————— 1. INTRODUCTION start of a new field of study which soon began to grow exponentially [6]. From that point, many of those re- ryopreservation is the fact of freezing cells or tissues C searches were performed in order to learn a way of freez- to preserve them in the future. Did you know that Darth ing sperm cell, so that in vitro fertilization became possi- Vader used it with Han Solo in one of the Star Wars mov- ble. ies? This method drags many years of researches related to the thermodynamic process of freezing and is based on In 1953 Jerome K. Sherman was successful in freezing some organisms like tardigrades. Moreover, it can be ex- and thawing human sperm, and on the same year he tended to many fields such as plants storage, surgery or founded the first sperm bank. Despite all of the discover- in vitro fecundation. ies taken place in the XX century, it was not until 1964 that the term cryobiology was invented and it was de- fined as a science.
    [Show full text]
  • Long-Term Cryopreservation and Revival of Tissue-Engineered Skeletal Muscle
    TISSUE ENGINEERING: Part A Volume 00, Number 00, 2018 ª Mary Ann Liebert, Inc. DOI: 10.1089/ten.tea.2018.0202 Long-Term Cryopreservation and Revival of Tissue-Engineered Skeletal Muscle Lauren Grant, MS,1,2,* Ritu Raman, PhD,3,* Caroline Cvetkovic, PhD,1,2,{ Meghan C. Ferrall-Fairbanks, PhD,4,{ Gelson J. Pagan-Diaz, MS,1,2 Pierce Hadley, BS,1 Eunkyung Ko, MS,1,2 Manu O. Platt, PhD,4 and Rashid Bashir, PhD1,2,5 Tissue-engineered skeletal muscle plays an important role not only in the field of regenerative medicine but also in emerging areas such as soft robotics, organ-on-a-chip disease models, and drug testing. However, further expansion of the applications of tissue-engineered skeletal muscle models requires a suitable method for their long-term storage and shipment. Cryopreservation has long been the standard for cell storage, but the freezing of three-dimensional tissues is accompanied by many complications due to heat and mass transfer limitations. In this study, we used a tissue-engineered skeletal muscle bioactuator as a model to characterize the effects of freezing on muscle viability, gene expression, myotube structure, and force generation. We optimized the protocol for cryopreservation by comparing outcomes when tissue was frozen undifferentiated and differentiated. Our opti- mized protocol, in which skeletal muscle was frozen undifferentiated, not only maintained cell viability but also led to a three-fold increase in force production compared to unfrozen muscle. Furthermore, we enhanced muscle lifetime through inhibition of cysteine proteases. The reported timeline for skeletal muscle tissue fabrication, freezing, revival, and long-term culture not only promotes a more streamlined fabrication process but also enables multisite collaborative research efforts through the shipment of preformed skeletal muscle constructs.
    [Show full text]