Thermally Reversible Hydrogel Sheets for Adult Stem Cell Culture

2010 ◽  
Author(s):  
Sreedhar Thirumala ◽  
Ram V. Devireddy
Keyword(s):  
Stem Cell ◽  
Cell Sheet ◽  
Adult Stem Cell ◽  
Collagen Type I ◽  
Type I ◽  
Non Invasive ◽  
Bovine Collagen ◽  
Invasive Method ◽  
Water Salt ◽  
Coated Surface

A simple non-invasive method of cell retrieval using Methylcellulose (MC) coated Tissue Culture Poly-Styrone (TCPS) dishes was developed to allow the creation of single and multilayered adult stem cell sheet constructs while preserving the cell-cell and the cell–extracellular matrices. The optimal combination of MC-water-salt was found to be 12 to 14% of MC (mol. wt. of 15,000) in water with 0.5x PBS (∼150 mOsm). This solution exhibited a gel formation temperature of ∼32°C. The addition (evenly spread) of 200μl of 2mg/ml bovine collagen type -I (pH adjusted to 7.5) over the MC coated surface at 37°C, significantly improved the adult stem cell (ASC) adhesion and proliferation on the hydrogel system.

scholarly journals Laminin-521 Promotes Rat Bone Marrow Mesenchymal Stem Cell Sheet Formation on Light-Induced Cell Sheet Technology

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Zhiwei Jiang ◽  
Yue Xi ◽  
Kaichen Lai ◽  
Ying Wang ◽  
Huiming Wang ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Cell Sheet ◽  
Sheet Thickness ◽  
Collagen Type I ◽  
Type I ◽  
Cell Sheets ◽  
Cell Sheet Technology ◽  
Rat Bone

Rat bone marrow mesenchymal stem cell sheets (rBMSC sheets) are attractive for cell-based tissue engineering. However, methods of culturing rBMSC sheets are critically limited. In order to obtain intact rBMSC sheets, a light-induced cell sheet method was used in this study. TiO2 nanodot films were coated with (TL) or without (TN) laminin-521. We investigated the effects of laminin-521 on rBMSCs during cell sheet culturing. The fabricated rBMSC sheets were subsequently assessed to study cell sheet viability, reattachment ability, cell sheet thickness, collagen type I deposition, and multilineage potential. The results showed that laminin-521 could promote the formation of rBMSC sheets with good viability under hyperconfluent conditions. Cell sheet thickness increased from an initial 26.7 ± 1.5 μm (day 5) up to 47.7 ± 3.0 μm (day 10). Moreover, rBMSC sheets maintained their potential of osteogenic, adipogenic, and chondrogenic differentiation. This study provides a new strategy to obtain rBMSC sheets using light-induced cell sheet technology.

Mechanical Stimulation Increases Collagen Type I and Collagen Type III Gene Expression of Stem Cell–Collagen Sponge Constructs for Patellar Tendon Repair

2007 ◽  
Vol 13 (6) ◽  
pp. 1219-1226 ◽  
Author(s):  
Natalia Juncosa-Melvin ◽  
Karl S. Matlin ◽  
Robert W. Holdcraft ◽  
Victor S. Nirmalanandhan ◽  
David L. Butler
Keyword(s):  
Gene Expression ◽  
Stem Cell ◽  
Patellar Tendon ◽  
Mechanical Stimulation ◽  
Collagen Type ◽  
Tendon Repair ◽  
Collagen Type I ◽  
Collagen Sponge ◽  
Type I ◽  
Collagen Type Iii

scholarly journals Optimizing growth factor induction of tenogenesis in three-dimensional culture of mesenchymal stem cells

2019 ◽  
Vol 10 ◽  
pp. 204173141984877 ◽  
Author(s):  
Ibtesam Rajpar ◽  
Jennifer G Barrett
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Growth Factors ◽  
Tendon Healing ◽  
Collagen Type I ◽  
Marker Genes ◽  
Type I ◽  
Cell Alignment ◽  
Tgf Β1

Adult tissue stem cells have shown promise for the treatment of debilitating tendon injuries. However, few comparisons of stem cells from different tissue sources have been made to determine the optimum stem cell source for treating tendon. Moreover, it is likely that the application of tenogenic growth factors will improve tendon stem cell treatments further, and a comprehensive comparison of a number of growth factors is needed. Thus far, different types of stem cells cannot be evaluated in a high-throughput manner. To this end, we have developed an approach to culture mesenchymal stem cells isolated from bone marrow in collagen type I hydrogels with tenogenic growth factors using economical, commercially available supplies. To optimize growth factors for this assay, FGF-2, TGF-β1, IGF-1, and/or BMP-12 were tested singly and in novel combinations of (1) BMP-12 and IGF-1, (2) TGF-β1 and IGF-1, and/or (3) BMP-12 and FGF-2 over 10 days. Our data suggest that BMP-12 supplementation alone results in the strongest expression of tendon marker genes, controlled contractility of constructs, a higher degree of cell alignment, and tendon-like tissue morphology. This easy-to-use benchtop assay can be used to screen novel sources of stem cells and cell lines for tissue engineering and tendon healing applications.

Analysis of Orientations of Collagen Fibers by Novel Fiber-Tracking Software

2003 ◽  
Vol 9 (6) ◽  
pp. 574-580 ◽  
Author(s):  
Jun Wu ◽  
Bartłomiej Rajwa ◽  
David L. Filmer ◽  
Christoph M. Hoffmann ◽  
Bo Yuan ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Collagen Type I ◽  
Type I ◽  
Collagen Gels ◽  
Microscopy Imaging ◽  
Bovine Collagen ◽  
Confocal Images ◽  
Tracing Algorithm ◽  
Highly Correlated ◽  
Processing Techniques

Recent evidence supports the notion that biological functions of extracellular matrix (ECM) are highly correlated to not only its composition but also its structure. This article integrates confocal microscopy imaging and image-processing techniques to analyze the microstructural properties of ECM. This report describes a two- and three-dimensional fiber middle-line tracing algorithm that may be used to quantify collagen fibril organization. We utilized computer simulation and statistical analysis to validate the developed algorithm. These algorithms were applied to confocal images of collagen gels made with reconstituted bovine collagen type I, to demonstrate the computation of orientations of individual fibers.

scholarly journals Refractory Metal Coated Alumina Foams as Support Material for Stem Cell and Fibroblasts Cultivation

Materials ◽  
2021 ◽  
Vol 14 (11) ◽  
pp. 2813
Author(s):  
Georg Hasemann ◽  
Ulf Betke ◽  
Manja Krüger ◽  
Heike Walles ◽  
Michael Scheffler
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Collagen Type I ◽  
Type I ◽  
Alumina Support ◽  
Average Cell Size ◽  
Average Cell ◽  
Cellular Behavior ◽  
Mediators Of Inflammation ◽  
Alumina Foam

Ceramics are widely used as implant materials; however, they are brittle and may emit particles when used in these applications. To overcome this disadvantage, alumina foams, which represent a 3D cellular structure comparable to that of human trabecular bone structures, were sputter coated with platinum, tantalum or titanium and modified with fibronectin or collagen type I, components of the extracellular matrix (ECM). To proof the cell material interaction, the unmodified and modified materials were cultured with (a) mesenchymal stem cells being a perfect indicator for biocompatibility and releasing important cytokines of the stem cell niche and (b) with fibroblasts characterized as mediators of inflammation and therefore an important cellular component of the foreign body reaction and inflammation after implantation. To optimize and compare the influence of metal surfaces on cellular behavior, planar glass substrates have been used. Identified biocompatible metal surface of platinum, titanium and tantalum were sputtered on ceramic foams modified with the above-mentioned ECM components to investigate cellular behavior in a 3D environment. The cellular alumina support was characterized with respect to its cellular/porous structure and niche accessibility and coating thickness of the refractory metals; the average cell size was 2.3 mm, the average size of the cell windows was 1.8 mm, and the total foam porosity was 91.4%. The Pt, Ti and Ta coatings were completely dense covering the entire alumina foam surface. The metals titanium and tantalum were colonized very well by the stem cells without a coating of ECM components, whereas the fibroblasts preferred components of the ECM on the alumina foam surface.

scholarly journals A simvastatin-releasing scaffold with periodontal ligament stem cell sheets for periodontal regeneration

2020 ◽  
Vol 18 ◽  
pp. 228080001990009
Author(s):  
Bingjiao Zhao ◽  
Jing Chen ◽  
Liru Zhao ◽  
Jiajia Deng ◽  
Qiang Li
Keyword(s):  
Osteogenic Differentiation ◽  
Periodontal Ligament ◽  
Cell Sheet ◽  
Periodontal Regeneration ◽  
Collagen Type I ◽  
Type I ◽  
Bovine Bone ◽  
Sustained Drug Release ◽  
Periodontal Ligament Stem Cells ◽  
Membrane Scaffold

Simvastatin (SIM) has been documented to induce the osteogenic differentiation of periodontal ligament stem cells (PDLSCs). To establish an efficient release system for periodontal regeneration, a polycaprolactone (PCL) membrane scaffold containing SIM was electrospun and evaluated. The obtained PCL–SIM membrane scaffold showed sustained release up to 28 days, without deleterious effect on proliferation of PDLSCs on the scaffolds. PDLSCs were seeded onto scaffolds and their osteogenic differentiation was evaluated. After 21 days, expressions of collagen type I, alkaline phosphatase and bone sialoprotein genes were significantly upregulated and mineralized matrix formation was increased on the PCL–SIM scaffolds compared with the PCL scaffolds. In a heterotopic periodontal regeneration model, a cell sheet–scaffold construct was assembled by placement of multilayers of PDLSC sheets on PCL or PCL–SIM scaffolds, and these were then placed between dentin and ceramic bovine bone for subcutaneous implantation in athymic mice. After 8 weeks, the PCL–SIM membrane showed formation of significantly more ectopic cementum-like mineral on the dentin surface. These findings demonstrated that the PCL–SIM membrane scaffold promotes cementum-like tissue formation by sustained drug release, suggesting the feasibility of its therapeutic use with PDLSC sheets to improve periodontal regeneration.

scholarly journals Effects of active and passive enrichment regimes on fecal glucocorticoid metabolite levels in captive Indian leopards (Panthera pardus fusca)

2021 ◽  
Author(s):  
Nirali Panchal ◽  
Chena Desai ◽  
Ratna Ghosal
Keyword(s):  
Environmental Enrichment ◽  
Medium Size ◽  
Type I ◽  
Health And Wellbeing ◽  
Non Invasive ◽  
Cage Size ◽  
Zoo Animals ◽  
Invasive Method ◽  
Free Ranging ◽  
Fecal Glucocorticoid

Environmental enrichment improves health and wellbeing of zoo animals. To test this hypothesis, we used Indian leopards, one of the popular zoo animals, as a model system to understand effects of active (interacting) and passive (noninteracting) enrichment elements on stress hormone levels of captive individuals. We included three enrichment categories, category ‘A’ (having both active: large size cage, and passive: controlled temperature, playback of forest sounds and sound proof glasses to filter visitors’ noise, enrichment elements), category ‘B’ (active enrichment type I, small size cage with air coolers), and category C (active enrichment type II, medium size cage without air coolers) for the leopards (n=14) housed in two Indian zoos. We standardized a non-invasive method to measure fecal glucocorticoid metabolite (fGCM) levels in captive leopards. The standardized fGCM assay was further validated by analysing samples from free-ranging leopards, as well. The fGCM levels (Mean±SE) were 10.45±2.01 and 0.95±0.003μg/g dry wt of feces in captive and free-ranging leopards, respectively. Our results demonstrated that fGCM levels of leopards in categories B and C were significantly (P<0.05) different from each other, thus, indicating cage size (an active enrichment element) as an important factor in influencing the physiology of the sampled animals. Overall, the findings of the study will contribute towards informing policies for management of the Indian leopards.

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