Direct cryopreservation of adherent cells on an elastic nanofiber sheet featuring a low glass-transition temperature

Batnyam, Onon; Suye, Shin‐ichiro; Fujita, Shin

doi:10.1039/c7ra10604a

Cited by 32 publications

(34 citation statements)

References 36 publications

(35 reference statements)

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“…Interestingly, after 7 days of 3D paper‐based cell culture, followed by freezing for 2 days at −80 °C, and finally thawing; experiments showed integrity and survival of cells (Figure S8b, Supporting Information). Obviously, and along with previously published reports on various scaffold‐based cell cryopreservations,8,9,27,28 paper‐based cryopreservation additionally overcomes the obstacles related to cryopreserving precultured (i.e., adherent) cells on 3D porous environments; namely intracellular crystallization, dehydration injury, mechanical ruptures, and uneven cooling during the freeze.…”

Section: Discussionmentioning

confidence: 88%

Paper‐Based Cell Cryopreservation

et al. 2020

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The continuous development of simple and practical cell cryopreservation methods is of great importance to a variety of sectors, especially when considering the efficient short‐ and long‐term storage of cells and their transportation. Although the overall success of such methods has been increased in recent years, there is still need for a unified platform that is highly suitable for efficient cryogenic storage of cells in addition to their easy‐to‐manage retrieval. Here, a paper‐based cell cryopreservation method as an alternative to conventional cryopreservation methods is presented. The method is space‐saving, cost‐effective, simple and easy to manage, and requires no additional fine‐tuning to conventional freezing and thawing procedures to yield comparable recovery of viable cells. It is shown that treating papers with fibronectin solution enhances the release of viable cells post thawing as compared to untreated paper platforms. Additionally, upon release, the remaining cells within the paper lead to the formation and growth of spheroid‐like structures. Moreover, it is demonstrated that the developed method works with paper‐based 3D cultures, where preformed 3D cultures can be efficiently cryopreserved.

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Section: Discussionmentioning

confidence: 88%

Paper‐Based Cell Cryopreservation

et al. 2020

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“…[ 64 ] A direct comparison of cell recovery and attachment on polymeric meshes varying in T g and fiber orientation revealed that cell recovery of adherent mouse C2C12 myoblasts could be enhanced using scaffolds composed of fibers of low‐ T g polymers oriented in a random instead of aligned manner. [ 65 ] Both characteristics enabled cells to remain (more) attached even upon dehydration during freezing. Scaffolds of polymers with a T g below the freezing temperature of the cryopreservation medium (approximately −20 °C for medium supplemented with 10% DMSO) remain more elastic.…”

Section: Cryopreservation Of Cardiac Samplesmentioning

confidence: 99%

“…Indeed, cell adhesions were found to persist after cryopreservation for random meshes, whereas cell detachment occurred especially parallel to the long fiber axis of aligned (and tensioned) fiber meshes. Scaffolds composed of low‐ T g polymers (of sufficiently low molecular weight), such as polyurethane (PU; T g −40 °C) [ 65 ] or PCL ( T g −60 °C), [ 63 ] or mixtures of low‐ and high‐ T g polymers, such as PCL/CS and SPCL meshes, are thus preferred over aligned meshes formed from high‐ T g polymers such as polystyrene (PS; T g 106 °C), [ 65 ] so that the mesh remains in a rubbery and deformable state while the cells shrink during freezing of the culture medium after which further cooling to storage temperatures below the T g induces an increase in stiffness and possibly crystallinity of the scaffold material.…”

Section: Cryopreservation Of Cardiac Samplesmentioning

confidence: 99%

“…[ 50 ] Also, cell retention during cryopreservation was shown to be negatively affected when polymer contraction was higher than cell shrinkage during the freezing phase of fiber scaffolds. Polymer temperature‐induced contraction is related to its crystallinity; [ 65 ] thus, it is crucial to select polymers with a T g lower than the preservation temperature.…”

Section: A Roadmap To Cte Construct Preservationmentioning

confidence: 99%

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A Roadmap to Cardiac Tissue‐Engineered Construct Preservation: Insights from Cells, Tissues, and Organs

et al. 2021

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Worldwide, over 26 million patients suffer from heart failure (HF). One strategy aspiring to prevent or even to reverse HF is based on the transplantation of cardiac tissue‐engineered (cTE) constructs. These patient‐specific constructs aim to closely resemble the native myocardium and, upon implantation on the diseased tissue, support and restore cardiac function, thereby preventing the development of HF. However, cTE constructs off‐the‐shelf availability in the clinical arena critically depends on the development of efficient preservation methodologies. Short‐ and long‐term preservation of cTE constructs would enable transportation and direct availability. Herein, currently available methods, from normothermic‐ to hypothermic‐ to cryopreservation, for the preservation of cardiomyocytes, whole‐heart, and regenerative materials are reviewed. A theoretical foundation and recommendations for future research on developing cTE construct specific preservation methods are provided. Current research suggests that vitrification can be a promising procedure to ensure long‐term cryopreservation of cTE constructs, despite the need of high doses of cytotoxic cryoprotective agents. Instead, short‐term cTE construct preservation can be achieved at normothermic or hypothermic temperatures by administration of protective additives. With further tuning of these promising methods, it is anticipated that cTE construct therapy can be brought one step closer to the patient.

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“…et al, 2012), electrospun-polyurethane nanofiber sheets (Batnyam et al, 2017), alginate-gelatin cryogel sponges (Katsen-Globa et al, 2014), and reticulated polyvinyl formal resins (Miyoshi et al, 2010). The results of these studies proved that the biocompatibility, mechanical support, and 3D porosity of scaffolds provide a suitable balance in creating a protective microenvironment for the cells during their cryopreservation.…”

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confidence: 98%

A Method to Efficiently Cryopreserve Mammalian Cells on Paper Platforms

Deliorman¹,

Sukumar²,

Alnemari³

et al. 2020

BIO-PROTOCOL

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This protocol describes a simple method to cryopreserve mammalian cells within filter papers as an alternative to conventional slow-freezing approach. The method involves treating paper fibers with fibronectin, using low concentrations of the cryoprotectant dimethyl sulfoxide (DMSO), and slow freezing cells to-80 °C at a 1 °C min-1 rate. In our method, the biocompatibility, large surface area, 3D porosity and fiber flexibility of the paper, in combination with the fibronectin treatment, yield recovery of cells comparable to conventional approaches, with no additional fine-tuning to freezing and thawing procedures. We expect that the paper-based cryopreservation method will bring several advantages to the field of preserving mammalian cells, including accommodation of a higher number of cells within a unit volume and no cell loss after release. The method requires a minimal storage space, where paper platforms with large areas can be rolled and/or folded and stored in stocks, and allows for efficient transportation/distribution of cells in an on-demand manner. Moreover, an additional feature of this method includes the formation and cryopreservation of cellular spheroids and 3D cell cultures.

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Direct cryopreservation of adherent cells on an elastic nanofiber sheet featuring a low glass-transition temperature

Abstract: Electrospun nanofibers, featured a lower glass-transition temperature than the freezing temperature and a loose mesh structure, allows the direct cryopreservation of adherent cells towards the investigation of cell-material composites.

Cited by 32 publications

References 36 publications

Paper‐Based Cell Cryopreservation

Paper‐Based Cell Cryopreservation

A Roadmap to Cardiac Tissue‐Engineered Construct Preservation: Insights from Cells, Tissues, and Organs

A Method to Efficiently Cryopreserve Mammalian Cells on Paper Platforms

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