Cell sheet tissue engineering for scaffold-free three-dimensional (3D) tissue reconstruction

Kim, Kyung-Sook; Bou-Ghannam, Sophia; Okano, Teruo

doi:10.1016/bs.mcb.2019.11.020

Cited by 17 publications

(21 citation statements)

References 87 publications

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“…Passage 5 hBMSCs were aliquoted in 20% FBS growth media 80 at 2.5 × 10 5 cells in 15 mL conical tubes for standard pellet cultures 22 , 61 , and seeded at 6.7 × 10 4 cells/cm 2 into 1.0 µm-diameter pore, 6-well cell culture inserts (Falcon, NE, USA) for monolayer cultures and into 35 mm diameter UpCell dishes (CellSeed, Tokyo, Japan) for cell sheet cultures based on previous methods for cell sheet fabrication 81 , 82 . For pellet fabrication, conical tubes were centrifuged at 500 × g for 10 min.…”

Section: Methodsmentioning

confidence: 99%

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

Thorp

Kim

Kondō

et al. 2020

Sci Rep

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Cell 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.

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Section: Methodsmentioning

confidence: 99%

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

Thorp

Kim

Kondō

et al. 2020

Sci Rep

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“…As a unique scaffold-free platform, cell sheet tissue engineering, pioneered by Okano et al. uses temperature-responsive cultureware to produce 3D cell sheet constructs that retain endogenous cellular environments, cell matrix, receptors, and adhesive proteins, permitting spontaneous sheet adhesion to biologic surfaces without supporting materials [ [22] , [23] , [24] , [25] , [26] , [27] ]. Previous work from our group has shown that 3D MSC sheets are capable of producing hyaline-like cartilage in vitro that directly adheres to cartilage surfaces [ 28 ].…”

Section: Introductionmentioning

confidence: 99%

Enhancing chondrogenic potential via mesenchymal stem cell sheet multilayering

Thorp

Kim

Bou-Ghannam

et al. 2021

Regenerative Therapy

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Advanced tissue engineering approaches for direct articular cartilage replacement in vivo employ mesenchymal stem cell (MSC) sources, exploiting innate chondrogenic potential to fabricate hyaline-like constructs in vitro within three-dimensional (3D) culture conditions. Cell sheet technology represents one such advanced 3D scaffold-free cell culture platform, and previous work has shown that 3D MSC sheets are capable of in vitro hyaline-like chondrogenic differentiation. The present study aims to build upon this understanding and elucidate the effects of an established cell sheet manipulation technique, cell sheet multilayering, on fabrication of MSC-derived hyaline-like cartilage 3D layered constructs in vitro. To achieve this goal, multilayered MSC sheets are prepared and assessed for structural and biochemical transitions throughout chondrogenesis. Results support MSC multilayering as a means of increasing construct thickness and 3D cellular interactions related to in vitro chondrogenesis, including N-cadherin, connexin 43, and integrin β-1. Data indicate that increasing construct thickness from 14 μm (1-layer construct) to 25 μm (2-layer construct) increases these cellular interactions and subsequent in vitro MSC chondrogenesis. However, a clear initial thickness threshold (33 μm - 3-layer construct) is evident that decreases the rate and extent of in vitro chondrogenesis, specifically chondrogenic gene expressions (Sox9, aggrecan, type II collagen) and sulfated proteoglycan accumulation in deposited extracellular matrix (ECM). Together, these data support the utility of cell sheet multilayering as a platform for tailoring construct thickness and subsequent MSC chondrogenesis for future articular cartilage regeneration applications.

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“…Dense membranes formed by the extracellular matrix and cells secreted by cells themselves effectively repair tissue damage. Based on the above advantages, cell membrane technology as a new cell transplantation method has been widely used in the field of regenerative medicine [ 111 , 112 ].…”

Section: Tissue Engineering Based Methodsmentioning

confidence: 99%

Recent trends in therapeutic strategies for repairing endometrial tissue in intrauterine adhesion

Ma¹,

Zhan

et al. 2021

Biomater Res

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Intrauterine adhesion (IUA) is a common gynaecological disease that develops from infection or trauma. IUA disease may seriously affect the physical and mental health of women of childbearing age, which may lead to symptoms such as hypomenorrhea or infertility. Presently, hysteroscopic transcervical resection of adhesion (TCRA) is the principal therapy for IUAs, although its function in preventing the recurrence of adhesion and preserving fertility is limited. Pharmaceuticals such as hormones and vasoactive agents and the placement of nondegradable stents are the most common postoperative adjuvant therapy methods. However, the repair of injured endometrium is relatively restricted due to the different anatomical structures of the endometrium. Recently, the treatment outcome of IUAs has improved with the advancement of hysteroscopic techniques. In particular, the application of bioactive scaffolds combined with tissue engineering technology has proven to have high therapeutic potential or endometrial repair in IUA treatment. Herein, this review has summarized past therapeutic strategies, including postoperative adjuvant therapy, cell or therapeutic molecular delivery therapy methods and bioactive scaffold-based tissue engineering methods. Therefore, this review presented the recent therapeutic strategies for repairing endometrium treatment and pointed out the issues of clinical concern to provide alternative methods for the management of IUAs.

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Cell sheet tissue engineering for scaffold-free three-dimensional (3D) tissue reconstruction

Cited by 17 publications

References 87 publications

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

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

Enhancing chondrogenic potential via mesenchymal stem cell sheet multilayering

Recent trends in therapeutic strategies for repairing endometrial tissue in intrauterine adhesion

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