Bulk poly(N-isopropylacrylamide) (PNIPAAm) thermoresponsive cell culture platform: toward a new horizon in cell sheet engineering

2019 ◽  
Vol 7 (6) ◽  
pp. 2277-2287 ◽  
Author(s):  
Andrew Choi ◽  
Kyoung Duck Seo ◽  
Hyungjun Yoon ◽  
Seon Jin Han ◽  
Dong Sung Kim
Keyword(s):  
Cell Culture ◽  
Cell Sheet ◽  
Cell Sheets ◽  
Cell Sheet Engineering

In contrast to the conventional ‘grafting’-based thermoresponsive cell culture platform, we first developed a bulk form of thermoresponsive cell culture platform for attaching/detaching diverse types and origins of the cell sheets in different shape.

scholarly journals Design of Temperature-Responsive Cell Culture Surfaces for Cell Sheet Engineering

2021 ◽  
Vol 2021 ◽  
pp. 1-15
Author(s):  
Y. Akiyama
Keyword(s):  
Cell Culture ◽  
Cell Attachment ◽  
Cell Sheet ◽  
Extensive Study ◽  
Cell Sheets ◽  
Cell Sheet Engineering ◽  
Temperature Responsive ◽  
Interdisciplinary Field ◽  
Culture Surface ◽  
Responsive Cell

Temperature-responsive cell culture surfaces, which modulate cell attachment/detachment characteristics with temperature, have been used to fabricate cell sheets. Extensive study on fabrication of cell sheet with the temperature-responsive cell culture surface, manipulation, and transplantation of the cell sheet has established the interdisciplinary field of cell sheet engineering, in which engineering, biological, and medical fields closely collaborate. Such collaboration has pioneered cell sheet engineering, making it a promising and attractive technology in tissue engineering and regenerative medicine. This review introduces concepts of cell sheet engineering, followed by designs for the fabrication of various types of temperature-responsive cell culture surfaces and technologies for cell sheet manipulation. The development of various methods for the fabrication of temperature-responsive cell culture surfaces was also summarized. The availability of cell sheet engineering for the treatment and regeneration of damaged human tissue has also been described, providing examples of the clinical application of cell sheet transplantation in humans.

scholarly journals Recent Advances in ROS-Responsive Cell Sheet Techniques for Tissue Engineering

2019 ◽  
Vol 20 (22) ◽  
pp. 5656 ◽  
Author(s):  
Min-Ah Koo ◽  
Mi Hee Lee ◽  
Jong-Chul Park
Keyword(s):  
Tissue Engineering ◽  
Cell Sheet ◽  
Cell Detachment ◽  
Oxygen Species ◽  
Cell Sheets ◽  
Cell Sheet Engineering ◽  
New Approach ◽  
Responsive Systems ◽  
Cell Based Therapy ◽  
Responsive Cell

Cell sheet engineering has evolved rapidly in recent years as a new approach for cell-based therapy. Cell sheet harvest technology is important for producing viable, transplantable cell sheets and applying them to tissue engineering. To date, most cell sheet studies use thermo-responsive systems to detach cell sheets. However, other approaches have been reported. This review provides the progress in cell sheet detachment techniques, particularly reactive oxygen species (ROS)-responsive strategies. Therefore, we present a comprehensive introduction to ROS, their application in regenerative medicine, and considerations on how to use ROS in cell detachment. The review also discusses current limitations and challenges for clarifying the mechanism of the ROS-responsive cell sheet detachment.

scholarly journals Cell Sheets Restore Secretory Function in Wounded Mouse Submandibular Glands

Cells ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 2645
Author(s):  
Harim T. dos Santos ◽  
Kyungsook Kim ◽  
Teruo Okano ◽  
Jean M. Camden ◽  
Gary A. Weisman ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Sheet ◽  
Early Time ◽  
Cell Polarization ◽  
Cell Sheets ◽  
Long Term Stability ◽  
Tissue Integrity ◽  
Tissue Organization ◽  
Post Surgery ◽  
Salivary Gland Regeneration

Thermoresponsive cell culture plates release cells as confluent living sheets in response to small changes in temperature, with recovered cell sheets retaining functional extracellular matrix proteins and tight junctions, both of which indicate formation of intact and functional tissue. Our recent studies demonstrated that cell sheets are highly effective in promoting mouse submandibular gland (SMG) cell differentiation and recovering tissue integrity. However, these studies were performed only at early time points and extension of the observation period is needed to investigate duration of the cell sheets. Thus, the goal of this study was to demonstrate that treatment of wounded mouse SMG with cell sheets is capable of increasing salivary epithelial integrity over extended time periods. The results indicate that cell sheets promote tissue organization as early as eight days after transplantation and that these effects endure through Day 20. Furthermore, cell sheet transplantation in wounded SMG induces a significant time-dependent enhancement of cell polarization, differentiation and ion transporter expression. Finally, this treatment restored saliva quantity to pre-wounding levels at both eight and twenty days post-surgery and significantly improved saliva quality at twenty days post-surgery. These data indicate that cell sheets engineered with thermoresponsive cell culture plates are useful for salivary gland regeneration and provide evidence for the long-term stability of cell sheets, thereby offering a potential new therapeutic strategy for treating hyposalivation.

scholarly journals Temperature-responsive intelligent interfaces for biomolecular separation and cell sheet engineering

2009 ◽  
Vol 6 (suppl_3) ◽  
Author(s):  
Kenichi Nagase ◽  
Jun Kobayashi ◽  
Teruo Okano
Keyword(s):  
Cell Sheet ◽  
Copolymer Composition ◽  
Cell Sheets ◽  
Chromatographic Separations ◽  
Cell Sheet Engineering ◽  
External Temperature ◽  
Cell Monolayers ◽  
Temperature Responsive ◽  
Confluent Cell ◽  
Dependent Cell

Temperature-responsive intelligent surfaces, prepared by the modification of an interface with poly( N -isopropylacrylamide) and its derivatives, have been used for biomedical applications. Such surfaces exhibit temperature-responsive hydrophilic/hydrophobic alterations with external temperature changes, which, in turn, result in thermally modulated interactions with biomolecules and cells. In this review, we focus on the application of these intelligent surfaces to chromatographic separation and cell cultures. Chromatographic separations using several types of intelligent surfaces are mentioned briefly, and various effects related to the separation of bioactive compounds are discussed, including wettability, copolymer composition and graft polymer architecture. Similarly, we also summarize temperature-responsive cell culture substrates that allow the recovery of confluent cell monolayers as contiguous living cell sheets for tissue-engineering applications. The key factors in temperature-dependent cell adhesion/detachment control are discussed from the viewpoint of grafting temperature-responsive polymers, and new methodologies for effective cell sheet culturing and the construction of thick tissues are summarized.

scholarly journals Scaffold-Free 3-D Cell Sheet Technique Bridges the Gap between 2-D Cell Culture and Animal Models

2019 ◽  
Vol 20 (19) ◽  
pp. 4926 ◽  
Author(s):  
Ayidah Alghuwainem ◽  
Alaa T. Alshareeda ◽  
Batla Alsowayan
Keyword(s):  
Tissue Engineering ◽  
Cell Culture ◽  
Animal Models ◽  
Tissue Repair ◽  
Cell Sheet ◽  
Cell Sheets ◽  
Systemic Injection ◽  
In Vivo Testing ◽  
D Cell

Various tissue engineering techniques have been created in research spanning two centuries, resulting in new opportunities for growing cells in culture and the creation of 3-D tissue-like constructs. These techniques are classified as scaffold-based and scaffold-free techniques. Cell sheet, as a scaffold-free technique, has attracted research interest in the context of drug discovery and tissue repair, because it provides more predictive data for in vivo testing. It is one of the most promising techniques and has the potential to treat degenerative tissues such as heart, kidneys, and liver. In this paper, we argue the advantages of cell sheets as a scaffold-free approach, compared to other techniques, including scaffold-based and scaffold-free techniques such as the classic systemic injection of cell suspension.

scholarly journals Phenotypic traits of mesenchymal stem cell sheets fabricated by temperature-responsive cell culture plate: structural characteristics of MSC sheets

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Mitsuyoshi Nakao ◽  
Kyungsook Kim ◽  
Kenichi Nagase ◽  
David W. Grainger ◽  
Hideko Kanazawa ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Cell Sheet ◽  
Cellular Structures ◽  
Phenotypic Traits ◽  
Cell Sheets ◽  
Temperature Responsive ◽  
Cell Sheet Technology ◽  
Responsive Cell

Abstract Background In most stem cell therapy strategies reported to date, stem cells are introduced to damaged tissue sites to repair and regenerate the original tissue structure and function. MSC therapeutic efficacies are inconsistent, largely attributed to transplanted MSC difficulties both in engrafting at tissue sites and in retaining their therapeutic functions from suspension formulations. MSC functional components, including cell adhesion and cell–cell junction proteins, and ECM that contribute to essential cellular therapeutic effects, are damaged or removed by proteolytic enzymes used in stem cell harvesting strategies from culture. To overcome these limitations, methods to harvest and transplant cells without disrupting critical stem cell functions are required. Cell sheet technology, exploiting temperature-responsive cell culture surfaces, permits cell harvest without cell protein damage. This study is focused on phenotypic traits of MSC sheets structurally and functionally to understand therapeutic benefits of cell sheets. Methods/results This study verified cleaved cellular proteins (vinculin, fibronectin, laminin, integrin β-1, and connexin 43) and increased apoptotic cell death produced under standard trypsin harvesting treatment in a time-dependent manner. However, MSC sheets produced without trypsin using only temperature-controlled sheet harvest from culture plastic exhibited intact cellular structures. Also, MSCs harvested using enzymatic treatment (i.e., chemical disruption) showed higher pYAP expression compared to MSC sheets. Conclusion Retention of cellular structures such as ECM, cell–cell junctions, and cell–ECM junctions is correlated with human umbilical cord mesenchymal stem cell (hUC-MSC) survival after detachment from cell culture surfaces. Retaining these proteins intact in MSC cultures using cell sheet technology is proposed to enhance stem cell survival and their function in stem cell-based therapy.

Human Mesenchymal Stem Cell Sheets Improve Uterine Incision Repair in a Rodent Hysterotomy Model

2020 ◽  
Author(s):  
Goro Kuramoto ◽  
Ibrahim A. Hammad ◽  
Brett D. Einerson ◽  
Amanda A. Allshouse ◽  
Michelle Debbink ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Umbilical Cord ◽  
Cell Sheet ◽  
Control Group ◽  
Human Umbilical Cord ◽  
Cell Sheets ◽  
Uterine Scar ◽  
Mesenchymal Stem Cell Sheet

Objective The study aimed to assess the feasibility of creating and transplanting human umbilical cord mesenchymal stem cell sheets applied to a rat model of hysterotomy, and additionally to determine benefits of human umbilical cord mesenchymal stem cell sheet transplantation in reducing uterine fibrosis and scarring. Study Design Human umbilical cord mesenchymal stem cell sheets are generated by culturing human umbilical cord mesenchymal stem cells on thermo-responsive cell culture plates. The temperature-sensitive property of these culture dishes facilitates normal cell culture in a thin contiguous layer and allows for reliable recovery of intact stem cell sheets without use of destructive proteolytic enzymes.We developed a rat hysterotomy model using nude rats. The rat uterus has two distinct horns: one horn provided a control/untreated scarring site, while the second horn was the cell sheet transplantation site.On day 14 following surgery, complete uteri were harvested and subjected to histologic evaluations of all hysterotomy sites. Results The stem cell sheet culture process yielded human umbilical cord mesenchymal stem cell sheets with surface area of approximately 1 cm2.Mean myometrial thickness in the cell sheet-transplanted group was 274 µm compared with 191 µm in the control group (p = 0.02). Mean fibrotic surface area in the human umbilical cord mesenchymal stem cell sheet-transplanted group was 95,861 µm2 compared with 129,185 µm2 in the control group. Compared with control horn sites, cell sheet-transplanted horns exhibited significantly smaller fibrotic-to-normal myometrium ratios (0.18 vs. 0.27, respectively, p = 0.029). Mean number of fibroblasts in cell sheet-transplanted horns was significantly smaller than the control horns (483 vs. 716/mm2, respectively, p = 0.001). Conclusion Human umbilical cord mesenchymal stem cell sheet transplantation is feasible in a rat model of hysterotomy. Furthermore, use of stem cell sheets reduces fibroblast infiltration and uterine scar fibrotic tissue formation during hysterotomy healing, potentially mitigating risks of uterine scar formation. Key Points

scholarly journals Partial Decellularized Scaffold Combined with Autologous Nasal Epithelial Cell Sheet for Tracheal Tissue Engineering

2021 ◽  
Vol 22 (19) ◽  
pp. 10322
Author(s):  
Luong Huu Dang ◽  
Shih-Han Hung ◽  
Yuan Tseng ◽  
Ly Xuan Quang ◽  
Nhi Thao Ngoc Le ◽  
...  
Keyword(s):  
Epithelial Cell ◽  
Cell Sheet ◽  
Rabbit Model ◽  
Immunohistochemical Analysis ◽  
Airway Epithelial ◽  
Potential Solution ◽  
Cell Sheets ◽  
Cell Sheet Engineering ◽  
Nasal Epithelial Cell

Decellularization has emerged as a potential solution for tracheal replacement. As a fully decellularized graft failed to achieve its purposes, the de-epithelialization partial decellularization protocol appeared to be a promising approach for fabricating scaffolds with preserved mechanical properties and few immune rejection responses after transplantation. Nevertheless, a lack of appropriate concurrent epithelialization treatment can lead to luminal stenosis of the transplant and impede its eventual success. To improve re-epithelialization, autologous nasal epithelial cell sheets generated by our cell sheet engineering platform were utilized in this study under an in vivo rabbit model. The newly created cell sheets have an intact and transplantable appearance, with their specific characteristics of airway epithelial origin being highly expressed upon histological and immunohistochemical analysis. Subsequently, those cell sheets were incorporated with a partially decellularized tracheal graft for autograft transplantation under tracheal partial resection models. The preliminary results two months post operation demonstrated that the transplanted patches appeared to be wholly integrated into the host trachea with adequate healing of the luminal surface, which was confirmed via endoscopic and histologic evaluations. The satisfactory result of this hybrid scaffold protocol could serve as a potential solution for tracheal reconstructions in the future.

scholarly journals Recent Advances in Engineered Stem Cell-Derived Cell Sheets for Tissue Regeneration

Polymers ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 209 ◽  
Author(s):  
Hyunbum Kim ◽  
Yunhye Kim ◽  
Jihyun Park ◽  
Nathaniel Hwang ◽  
Yun Lee ◽  
...  
Keyword(s):  
Stem Cell ◽  
Tissue Regeneration ◽  
Proteolytic Enzymes ◽  
Intact Cell ◽  
Cell Sheet ◽  
Intercellular Junctions ◽  
Surface Proteins ◽  
Cell Sheets ◽  
Cell Sheet Engineering ◽  
Cell Based Therapy

The substantial progress made in the field of stem cell-based therapy has shown its significant potential applications for the regeneration of defective tissues and organs. Although previous studies have yielded promising results, several limitations remain and should be overcome for translating stem cell-based therapies to clinics. As a possible solution to current bottlenecks, cell sheet engineering (CSE) is an efficient scaffold-free method for harvesting intact cell sheets without the use of proteolytic enzymes, and may be able to accelerate the adoption of stem cell-based treatments for damaged tissues and organs regeneration. CSE uses a temperature-responsive polymer-immobilized surface to form unique, scaffold-free cell sheets composed of one or more cell layers maintained with important intercellular junctions, cell-secreted extracellular matrices, and other important cell surface proteins, which can be achieved by changing the surrounding temperature. These three-dimensional cell sheet-based tissues can be designed for use in clinical applications to target-specific tissue regeneration. This review will highlight the principles, progress, and clinical relevance of current approaches in the cell sheet-based technology, focusing on stem cell-based therapies for bone, periodontal, skin, and vascularized muscles.

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