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

Asakawa, Nahoko; Shimizu, Tatsuya; Tsuda, Yukiko; Sekiya, Sachiko; Sasagawa, Tadashi; Yamato, Masayuki; Fukai, Fumio; Okano, Teruo

doi:10.1016/j.biomaterials.2010.01.105

Cited by 223 publications

(149 citation statements)

References 25 publications

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“…In the herein presented work, it was shown that HUVECs, seeded over rBMSCs 4 days before the end of the osteogenic differentiation period, self-organized either in small round colonies or in elongated aggregates resembling a network. As these cocultured CSs were not cultured in vitro after stacking with a second osteogenic CSs, which could provide a 3D matrix for HUVECs reorganization, the formation of capillary-like structures, as seen in other works, 23,24,35 was not expected.…”

Section: Discussionmentioning

confidence: 90%

“…23,24,35 Sasagawa et al 24 seeded HUVECs between myoblast CSs, building five-layer constructs, to obtain in vitro a capillary-like network that anastomosed when implanted in nude rats, allowing the survival of the construct. Another work 23 showed that the positioning of endothelial cells within two fibroblast CS constructs was determinant for tubular formation and influences its lumen area in vitro. Thus, it is increasingly clear that stacking several CS results in a suitable 3D environment for endothelial cells to form a prevascular network in vitro.…”

Section: Discussionmentioning

confidence: 99%

“…The prevascularization of CS-based 3D constructs has nevertheless been the focus of recent works. [23][24][25] The combination of several myoblasts sheets, recovered from PIPPAam dishes, and human umbilical vein endothelial cells (HUVECs) using a CS stacking methodology resulted in the generation of a prevascularized cell-dense tissue. 24 This work is a proof-of-concept on how CS engineering has the potential to produce 3D prevascularized tissues.…”

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confidence: 99%

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Endothelial cells enhance the in vivo bone-forming ability of osteogenic cell sheets

Pirraco

Iwata

Yoshida

et al. 2014

Laboratory Investigation

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Addressing the problem of vascularization is of vital importance when engineering three-dimensional (3D) tissues. Endothelial cells are increasingly used in tissue-engineered constructs to obtain prevascularization and to enhance in vivo neovascularization. Rat bone marrow stromal cells were cultured in thermoresponsive dishes under osteogenic conditions with human umbilical vein endothelial cells (HUVECs) to obtain homotypic or heterotypic cell sheets (CSs). Cells were retrieved as sheets from the dishes after incubation at 20 1C. Monoculture osteogenic CSs were stacked on top of homotypic or heterotypic CSs, and subcutaneously implanted in the dorsal flap of nude mice for 7 days. The implants showed mineralized tissue formation under both conditions. Transplanted osteogenic cells were found at the new tissue site, demonstrating CS bone-inductive effect. Perfused vessels, positive for human CD31, confirmed the contribution of HUVECs for the neovascularization of coculture CS constructs. Furthermore, calcium quantification and expression of osteocalcin and osterix genes were higher for the CS constructs, with HUVECs demonstrating the more robust osteogenic potential of these constructs. This work demonstrates the potential of using endothelial cells, combined with osteogenic CSs, to increase the in vivo vascularization of CS-based 3D constructs for bone tissue engineering purposes.

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Section: Discussionmentioning

confidence: 90%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Endothelial cells enhance the in vivo bone-forming ability of osteogenic cell sheets

Pirraco

Iwata

Yoshida

et al. 2014

Laboratory Investigation

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“…In the present investigation, the scaffoldless cell sheet technology, originally developed to fabricate rat mesenchymal and murine cardiac progenitor cell sheets [4,7], was adopted for the first time to fabricate human cardiac engineered tissues to emulate the natural hCPC environmental architecture and to minimize the stress caused by conventional manipulations. Indeed, engineered hCPCs retained their Sca-1 pos undifferentiated phenotype, but dramatically reduced their growth rate; in fact, genes involved in cell cycling were downregulated and very few apoptotic events occurred within 7 days after sheet fabrication.…”

Section: Discussionmentioning

confidence: 99%

“…To this aim, progenitor cells can be appropriately grown on polymeric scaffolds [2,3] or, alternatively, thermo-responsive surfaces [4][5][6]. Following the latter procedure, scaffoldless tissue sheets fabricated using clonally expanded stem cell antigen-1 positive (Sca-1 pos ) progenitor cells (CPCs) isolated from murine hearts, prevascularized [7] and transplanted into hearts with experimental myocardial infarction have shown to prevent cardiac Author contributions: G.F. and S.P. : human cell extraction, culture, and cell sheet characterization; F.P., G.N., E.M., and A.Z.…”

Section: Introductionmentioning

confidence: 99%

Human Cardiac Progenitor Cell Grafts as Unrestricted Source of Supernumerary Cardiac Cells in Healthy Murine Hearts

et al. 2011

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Human heart harbors a population of resident progenitor cells that can be isolated by stem cell antigen-1 antibody and expanded in culture. These cells can differentiate into cardiomyocytes in vitro and contribute to cardiac regeneration in vivo. However, when directly injected as single cell suspension, less than 1%-5% survive and differentiate. Among the major causes of this failure are the distressing protocols used to culture in vitro and implant progenitor cells into damaged hearts. Human cardiac progenitors obtained from the auricles of patients were cultured as scaffoldless engineered tissues fabricated using temperature-responsive surfaces. In the engineered tissue, progenitor cells established proper three-dimensional intercellular relationships and were embedded in self-produced extracellular matrix preserving their phenotype and multipotency in the absence of significant apoptosis. After engineered tissues were leant on visceral pericardium, a number of cells migrated into the murine myocardium and in the vascular walls, where they integrated in the respective textures. The study demonstrates the suitability of such an approach to deliver stem cells to the myocardium. Interestingly, the successful delivery of cells in murine healthy hearts suggests that myocardium displays a continued cell cupidity that is strictly regulated by the limited release of progenitor cells by the adopted source. When an unregulated cell source is added to the system, cells are delivered to the myocardium. The exploitation of this novel concept may pave the way to the setup of new protocols in cardiac cell therapy.

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Thermoresponsive Cell Culture Surfaces Designed for Cell‐Sheet‐Based Tissue Engineering and Regenerative Medicine

Kobayashi

Okano

2013

Biomaterials Surface Science

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

Cited by 223 publications

References 25 publications

Endothelial cells enhance the in vivo bone-forming ability of osteogenic cell sheets

Endothelial cells enhance the in vivo bone-forming ability of osteogenic cell sheets

Human Cardiac Progenitor Cell Grafts as Unrestricted Source of Supernumerary Cardiac Cells in Healthy Murine Hearts

Thermoresponsive Cell Culture Surfaces Designed for Cell‐Sheet‐Based Tissue Engineering and Regenerative Medicine

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