Multi-layered in vitro 3D-bone model via combination of osteogenic cell sheets with electrospun membrane interlayer

2021 ◽  
pp. 088532822110278
Author(s):  
Atakan Tevlek ◽  
Halil Murat Aydin
Keyword(s):  
Cell Sheet ◽  
Three Dimensional ◽  
Osteogenic Cell ◽  
Electrospun Membrane ◽  
Cell Sheets ◽  
Cell Sheet Engineering ◽  
Sulfated Glycosaminoglycan ◽  
Sgag Content ◽  
In Vitro Performance

In this study, it was aimed to present an approach for the development of multi-layered tissue engineering constructs by using cell sheet engineering. Briefly, MC3T3-E1 mouse pre-osteoblast cells were cultured in temperature-responsive plates (Nunc Upcell®) in the presence of osteogenic medium and the resulting cell sheets were laminated with electrospun poly(L-lactic acid) (PLLA) membranes to obtain viable three-dimensional, thick constructs. The constructs prepared without PLLA membranes were used as control. The cell viability and death in the resulting structures were investigated by microscopic and colorimetric methods. The in vitro performance of the structures was discussed comparatively. Alkaline phosphatase (ALP) activity, collagen and sulfated glycosaminoglycan (sGAG) content values were calculated. The presented approach shows potential for engineering applications of complex tissues with at least two or more microenvironments such as osteochondral, corneal or vascular tissues.

scholarly journals In Vitro Comparison of 2D-Cell Culture and 3D-Cell Sheets of Scleraxis-Programmed Bone Marrow Derived Mesenchymal Stem Cells to Primary Tendon Stem/Progenitor Cells for Tendon Repair

2018 ◽  
Vol 19 (8) ◽  
pp. 2272 ◽  
Author(s):  
Chi-Fen Hsieh ◽  
Zexing Yan ◽  
Ricarda Schumann ◽  
Stefan Milz ◽  
Christian Pfeifer ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Sheet ◽  
Tendon Repair ◽  
Three Dimensional ◽  
Cell Types ◽  
Tendon Tissue ◽  
Cell Sheets ◽  
Tendon Tissue Engineering ◽  
Dimensional Cell

The poor and slow healing capacity of tendons requires novel strategies to speed up the tendon repair process. Hence, new and promising developments in tendon tissue engineering have become increasingly relevant. Previously, we have established a tendon progenitor cell line via ectopic expression of the tendon-related basic helix-loop-helix (bHLH) transcription factor Scleraxis (Scx) in human bone marrow mesenchymal stem cells (hMSC-Scx). The aim of this study was to directly compare the characteristics of hMSC-Scx cells to that of primary human tendon stem/progenitors cells (hTSPCs) via assessment of self-renewal and multipotency, gene marker expression profiling, in vitro wound healing assay and three-dimensional cell sheet formation. As expected, hTSPCs were more naive than hMSC-Scx cells because of higher clonogenicity, trilineage differentiation potential, and expression of stem cell markers, as well as higher mRNA levels of several gene factors associated with early tendon development. Interestingly, with regards to wound healing, both cell types demonstrate a comparable speed of scratch closure, as well as migratory velocity and distance in various migration experiments. In the three-dimensional cell sheet model, hMSC-Scx cells and hTSPCs form compact tendinous sheets as histological staining, and transmission electron microscopy shows spindle-shaped cells and collagen type I fibrils with similar average diameter size and distribution. Taken together, hTSPCs exceed hMSC-Scx cells in several characteristics, namely clonogenicity, multipotentiality, gene expression profile and rates of tendon-like sheet formation, whilst in three-dimensional cell sheets, both cell types have comparable in vitro healing potential and collagenous composition of their three-dimensional cell sheets, making both cell types a suitable cell source for tendon tissue engineering and healing.

Pre-vascularization of in vitro three-dimensional tissues created by cell sheet engineering

Biomaterials ◽  
2010 ◽  
Vol 31 (14) ◽  
pp. 3903-3909 ◽  
Author(s):  
Nahoko Asakawa ◽  
Tatsuya Shimizu ◽  
Yukiko Tsuda ◽  
Sachiko Sekiya ◽  
Tadashi Sasagawa ◽  
...  
Keyword(s):  
Cell Sheet ◽  
Three Dimensional ◽  
Cell Sheet Engineering

Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

1974 ◽  
Vol 32 ◽  
pp. 320-321
Author(s):  
J. P. Revel
Keyword(s):  
Developmental Stages ◽  
Three Dimensional ◽  
Cell Movement ◽  
Cellular Interactions ◽  
Serial Sections ◽  
Cell Sheets ◽  
Freeze Etching ◽  
Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

Rapid fabrication system for three-dimensional tissues using cell sheet engineering and centrifugation

2015 ◽  
Vol 103 (12) ◽  
pp. 3825-3833 ◽  
Author(s):  
Akiyuki Hasegawa ◽  
Yuji Haraguchi ◽  
Tatsuya Shimizu ◽  
Teruo Okano
Keyword(s):  
Cell Sheet ◽  
Three Dimensional ◽  
Cell Sheet Engineering ◽  
Rapid Fabrication

Abstract 020: Cell-sheet-based Bioengineered Human Cardiac Tissue Using Pluripotent Stem Cells For Heart Repair And Disease Models

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Katsuhisa Matsuura ◽  
Tatsuya Shimizu ◽  
Nobuhisa Hagiwara ◽  
Teruo Okano
Keyword(s):  
Cardiac Tissue ◽  
Subcutaneous Tissue ◽  
Cell Sheet ◽  
Three Dimensional ◽  
Embryoid Bodies ◽  
Cardiac Differentiation ◽  
Cardiac Cell ◽  
Cell Sheets ◽  
High Efficient ◽  
Human Cardiac Tissue

We have developed an original scaffold-free tissue engineering approach, “cell sheet engineering”, and this technology has been already applied to regenerative medicine of various organs including heart. As the bioengineered three-dimensional cardiac tissue is expected to not only function for repairing the broad injured heart but also to be the practicable heart tissue models, we have developed the cell sheet-based perfusable bioengineered three-dimensional cardiac tissue. Recently we have also developed the unique suspension cultivation system for the high-efficient cardiac differentiation of human iPS cells. Fourteen-day culture with the serial treatments of suitable growth factors and a small compound in this stirring system with the suitable dissolved oxygen concentration produced robust embryoid bodies that showed the spontaneous beating and were mainly composed of cardiomyocytes (~80%). When these differentiated cells were cultured on temperature-responsive culture dishes after the enzymatic dissociation, the spontaneous and synchronous beating was observed accompanied with the intracellular calcium influx all over the area even after cell were detached from culture dishes as cell sheets by lowering the culture temperature. The cardiac cell sheets were mainly composed of cardiomyocytes (~80%) and partially mural cells (~20%). Furthermore, extracellular action potential propagation was observed between cell sheets when two cardiac cell sheets were partially overlaid, and this propagation was inhibited by the treatment with some anti-arrhythmic drugs. When the triple layered cardiac tissue was transplanted onto the subcutaneous tissue of nude rats, the spontaneous pulsation was observed over 2 months and engrafted cardiomyocytes were vascularized with the host tissue-derived endothelial cells. These findings suggest that cardiac cell sheets formed by hiPSC-derived cardiomyocytes might have sufficient properties for the creation of thickened cardiac tissue. Now we are developing the vascularized thickened human cardiac tissue by the repeated layering of cardiac cell sheets on the artificial vascular bed in vitro.

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 Fabrication of hyaline-like cartilage constructs using mesenchymal stem cell sheets

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Hallie Thorp ◽  
Kyungsook Kim ◽  
Makoto Kondo ◽  
David W. Grainger ◽  
Teruo Okano
Keyword(s):  
Chondrogenic Differentiation ◽  
Cell Sheet ◽  
Cartilage Regeneration ◽  
Cell Sheets ◽  
Chondrogenic Potential ◽  
Cell Sheet Technology ◽  
Cell Functionality ◽  
Articular Cartilage Regeneration

AbstractCell and tissue engineering approaches for articular cartilage regeneration increasingly focus on mesenchymal stem cells (MSCs) as allogeneic cell sources, based on availability and innate chondrogenic potential. Many MSCs exhibit chondrogenic potential as three-dimensional (3D) cultures (i.e. pellets and seeded biomaterial scaffolds) in vitro; however, these constructs present engraftment, biocompatibility, and cell functionality limitations in vivo. Cell sheet technology maintains cell functionality as scaffold-free constructs while enabling direct cell transplantation from in vitro culture to targeted sites in vivo. The present study aims to develop transplantable hyaline-like cartilage constructs by stimulating MSC chondrogenic differentiation as cell sheets. To achieve this goal, 3D MSC sheets are prepared, exploiting spontaneous post-detachment cell sheet contraction, and chondrogenically induced. Results support 3D MSC sheets’ chondrogenic differentiation to hyaline cartilage in vitro via post-contraction cytoskeletal reorganization and structural transformations. These 3D cell sheets’ initial thickness and cellular densities may also modulate MSC-derived chondrocyte hypertrophy in vitro. Furthermore, chondrogenically differentiated cell sheets adhere directly to cartilage surfaces via retention of adhesion molecules while maintaining the cell sheets’ characteristics. Together, these data support the utility of cell sheet technology for fabricating scaffold-free, hyaline-like cartilage constructs from MSCs for future transplantable articular cartilage regeneration therapies.

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 A Concise Review on the Use of Mesenchymal Stem Cells in Cell Sheet-Based Tissue Engineering with Special Emphasis on Bone Tissue Regeneration

2017 ◽  
Vol 2017 ◽  
pp. 1-13 ◽  
Author(s):  
A. Cagdas Yorukoglu ◽  
A. Esat Kiter ◽  
Semih Akkaya ◽  
N. Lale Satiroglu-Tufan ◽  
A. Cevik Tufan
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Ethical Issues ◽  
Cell Sheet ◽  
Bone Tissue Regeneration ◽  
Differentiation Potential ◽  
Cell Sheet Engineering ◽  
Application Techniques

The integration of stem cell technology and cell sheet engineering improved the potential use of cell sheet products in regenerative medicine. This review will discuss the use of mesenchymal stem cells (MSCs) in cell sheet-based tissue engineering. Besides their adhesiveness to plastic surfaces and their extensive differentiation potential in vitro, MSCs are easily accessible, expandable in vitro with acceptable genomic stability, and few ethical issues. With all these advantages, they are extremely well suited for cell sheet-based tissue engineering. This review will focus on the use of MSC sheets in osteogenic tissue engineering. Potential application techniques with or without scaffolds and/or grafts will be discussed. Finally, the importance of osteogenic induction of these MSC sheets in orthopaedic applications will be demonstrated.

Fabrication of functional three-dimensional tissues by stacking cell sheets in vitro

2012 ◽  
Vol 7 (5) ◽  
pp. 850-858 ◽  
Author(s):  
Yuji Haraguchi ◽  
Tatsuya Shimizu ◽  
Tadashi Sasagawa ◽  
Hidekazu Sekine ◽  
Katsuhisa Sakaguchi ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Cell Sheets
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