Engineered mesenchymal stem-cell-sheets patches prevents postoperative pancreatic leakage in a rat model

Kim, Seong-Ryong; Yi, Hye-Jin; Lee, Yu Na; Park, Ji Yoon; Hoffman, Robert M.; Okano, Teruo; Shim, In Kyong; Kim, Song Cheol

doi:10.1038/s41598-017-18490-9

Cited by 20 publications

(31 citation statements)

References 43 publications

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“…The Endothelial Progenitor Cells (EPC) treatment strategy take advantages of preserving or regenerating tissue to develop towards clinical application [1][2][3]. However, the disadvantage of single cells transplantation related aggregation and necrosis make the the feasibility and effectiveness of the approach remain insu cient and di cult in clinical trials [4,5].…”

Section: Introductionmentioning

confidence: 99%

Fabrication and Paracrine Effect Research of Bone Marrow-Derived Endothelial Progenitor Cell Sheet

Xue¹,

Yin²,

Bai³

et al. 2020

Preprint

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Background: The release of a wide array of endothelial progenitor cell (EPC) sheet-secreted paracrine factors is central to the mechanism by which these cells contribute to tissue repair. The purpose of this study was to fabricate BM-EPC sheet and conduct preliminary investigation on the paracrine effect of SDF-1ɑ about the role of the stromal cell-derived factor-1α (SDF-1α)/CXCR4 axis in the form of tubular structures from BM-EPCs sheet.Methods: EPCs derived from rat bone marrow (BM-EPC) were identified and isolated by the cell-surface markers CD34/CD133/VE-cadherin/KDR using flow cytometry, as well as by dual affinity for acLDL and UEA-1. Single-cell suspensions were seeded on temperature-responsive cell culture dishes. After 7 days of incubation, the BM-EPC cells were easily harvested as cell sheets, and a series of biochemical experiments were performed in vitro. The expression levels of SDF-1α/CXCR4 axis-associated genes and proteins were examined using RT‑qPCR and western blot analysis, and enzyme-linked immunosorbent assay (ELISA) was applied to determine the concentrations of VEGF, EGF and SDF-1α in the cell culture medium.Results: The BM-EPC cell sheets were successfully harvested. Moreover, BM-EPC cell sheets have superior proliferation and tube formation activity when compare with single cell suspension. When capillary-like tube were formed from EPCs sheets, the releasing of paracrine factors such as VEGF, EGF, and SDF-1ɑ were increasing. To drive the tube formation of BM-EPCs sheets, our mechanism research showed that the activation of PI3K/AKT/eNOS pathway since the phosphorylation of protein CXCR, PI3K, AKT and eNOS were increasing. Conclusion: BM-EPC cell sheets have superior proliferation and tube formation activity that can be used for tissue repair. The strong ability to secrete paracrine factors was be related to the SDF-1α/CXCR4 axis through PI3K/AKT/eNOS pathway.

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Section: Introductionmentioning

confidence: 99%

Fabrication and Paracrine Effect Research of Bone Marrow-Derived Endothelial Progenitor Cell Sheet

Xue¹,

Yin²,

Bai³

et al. 2020

Preprint

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“…Previously described cell-assembled systems for different tissues and organs may therefore take advantage of the use of our approach to accelerate the time required for ex vivo culture 33 – 39 . In a simple setting, our technology can be combined with the use of temperature-responsive plates, allowing the detachment of cell sheets by a temperature switch as already tested for macromolecular “crowders” 36 , 40 , 41 .…”

Section: Discussionmentioning

confidence: 99%

Enhancement of collagen deposition and cross-linking by coupling lysyl oxidase with bone morphogenetic protein-1 and its application in tissue engineering

Rosell-García

Rodrı́guez-Pascual

2018

Sci Rep

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Cultured cell-derived extracellular matrices (ECM)-based biomaterials exploit the inherent capacity of cells to create highly sophisticated supramolecular assemblies. However, standard cell culture conditions are far from ideal given the fact that the diluted microenvironment does not favor the production of ECM components, a circumstance particularly relevant for collagen. An incomplete conversion of procollagen by C-proteinase/bone morphogenetic protein 1 (BMP1) has been proposed to severely limit in vitro collagen deposition. BMP1 also catalyzes the proteolytic activation of the precursor of the collagen cross-linking enzyme, lysyl oxidase (LOX) to yield the active form, suggesting a deficit in cross-linking activity under standard conditions. We hypothesized that the implementation of fibroblast cultures with LOX and BMP1 may be an effective way to increase collagen deposition. To test it, we have generated stable cell lines overexpressing LOX and BMP1 and studied the effect of supernatants enriched in LOX and BMP1 on collagen synthesis and deposition from fibroblasts. Herein, we demonstrate that the supplementation with LOX and BMP1 strongly increased the deposition of collagen onto the insoluble matrix at the expense of the soluble fraction in the extracellular medium. Using decellularization protocols, we also show that fibroblast-derived matrices regulate adipogenic and osteogenic differentiation of human mesenchymal stem cells (MSC), and this effect was modulated by LOX/BMP1. Collectively, these data demonstrate that we have developed a convenient protocol to enhance the capacity of in vitro cell cultures to deposit collagen in the ECM, representing this approach a promising technology for application in tissue engineering.

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“…Over 15 years of clinical data support enhanced therapeutic properties for autologous‐sourced cell sheets through increased cell retention at target tissues (Ebihara et al, 2012; Iwata et al, 2018; Kanzaki, Takagi, Washio, Kokubo, & Yamato, 2017; Nishida et al, 2004; Ohki et al, 2012; Sawa et al, 2015; Yamaguchi et al, 2017; Yamamoto et al, 2017). Recently, several preclinical studies have also reported MSC sheet use in wound healing, heart, and pancreas regeneration (Kato et al, 2015; Kim et al, 2018; Miyahara et al, 2006). However, for reliable clinical performance and regulatory acceptance, standardized MSC sheet fabrication methods yielding reproducible cell‐based products with quality control specifications must first be developed.…”

Section: Introductionmentioning

confidence: 99%

Stable cell adhesion affects mesenchymal stem cell sheet fabrication: Effects of fetal bovine serum and human platelet lysate

Kim

Thorp

Bou-Ghannam

et al. 2020

J Tissue Eng Regen Med

Self Cite

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Cell sheet technology exploits temperature responsive cell culture dishes (TRCDs) as versatile cell harvesting methods to yield contiguous cell monolayers robustly held together by cell-cell junctions, receptors, and endogenous extracellular matrix. More than 15 years of clinical data using autologous-sourced cell sheets demonstrate enhanced therapeutic properties through increased cell retention at target tissue sites. Recently, several preclinical studies have also been reported using mesenchymal stem cell (MSC) sheets in wound healing, cardiac ischemia therapies, and pancreatic regeneration. However, optimized MSC sheet fabrication conditions have not yet been reported. In this study, we identified specific conditions for reliable human MSC sheet fabrication by comparing cell growth media supplements (fetal bovine serum [FBS] and human platelet lysate [hPL]). Human umbilical cord-derived MSCs cultured in FBS and hPL exhibit different actin cytoskeletal structures related to their cell morphologies and adhesion. MSCs cultured in FBS media showed stable cell adhesion on TRCDs with flattened cell shapes and aligned actin cytoskeletal structure. This stable cell adhesion enables production of consistent MSC cell sheets, with controlled cell sheet detachment. Conversely, cell sheet fabrication in hPL media exhibits poor reproducibility being more sensitive to temperature-and culture timeinduced release due to weak cell adhesion. These findings suggest that stable MSC adhesion to TRCDs is important to reliable MSC sheet fabrication methods and that MSC growth media supplementation directly affects cell adhesion during culture. K E Y W O R D S cell adhesion, cell morphology, cell therapy, cytokine secretion, cytoskeleton structure, fetal bovine serum, human platelet lysate

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Engineered mesenchymal stem-cell-sheets patches prevents postoperative pancreatic leakage in a rat model

Cited by 20 publications

References 43 publications

Fabrication and Paracrine Effect Research of Bone Marrow-Derived Endothelial Progenitor Cell Sheet

Fabrication and Paracrine Effect Research of Bone Marrow-Derived Endothelial Progenitor Cell Sheet

Enhancement of collagen deposition and cross-linking by coupling lysyl oxidase with bone morphogenetic protein-1 and its application in tissue engineering

Stable cell adhesion affects mesenchymal stem cell sheet fabrication: Effects of fetal bovine serum and human platelet lysate

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