scholarly journals 3D arrays of microcages by two-photon lithography for spatial organization of living cells

Lab on a Chip ◽  
2019 ◽  
Vol 19 (5) ◽  
pp. 875-884 ◽  
Author(s):  
Florian Larramendy ◽  
Shotaro Yoshida ◽  
Daniela Maier ◽  
Zoltan Fekete ◽  
Shoji Takeuchi ◽  
...  
Keyword(s):  
Neuronal Networks ◽  
Spatial Organization ◽  
Three Dimensional ◽  
Living Cells ◽  
Artificial Organs ◽  
Two Photon ◽  
3D Network

This paper addresses a nanoengineering approach to create a fully three-dimensional (3D) network of living cells, providing an advanced solution to in vitro studies on either neuronal networks or artificial organs.

scholarly journals The lighthouse at the end of the chromosome*

F1000Research ◽  
2015 ◽  
Vol 4 ◽  
pp. 1427 ◽  
Author(s):  
Yahya Benslimane ◽  
Lea Harrington
Keyword(s):  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Spatial Organization ◽  
Three Dimensional ◽  
Time Lapse ◽  
Living Cells ◽  
Telomere Elongation ◽  
Cellular Processes ◽  
Time Lapse Microscopy

Fluorescence microscopy can be used to assess the dynamic localization and intensity of single entities in vitro or in living cells. It has been applied with aplomb to many different cellular processes and has significantly enlightened our understanding of the heterogeneity and complexity of biological systems. Recently, high-resolution fluorescence microscopy has been brought to bear on telomeres, leading to new insights into telomere spatial organization and accessibility, and into the mechanistic nuances of telomere elongation. We provide a snapshot of some of these recent advances with a focus on mammalian systems, and show how three-dimensional, time-lapse microscopy and single-molecule fluorescence shine a new light on the end of the chromosome.

Associated Anisotropy of intrinsic NAD(P)H for monitoring changes in the metabolic activities of breast cancer cells in three-dimensional collagen matrix

2021 ◽  
Author(s):  
Anh Cong ◽  
Rafaela M. L. Pimenta ◽  
Jon Holy ◽  
Ahmed A Heikal
Keyword(s):  
Breast Cancer ◽  
Cell Culture ◽  
Breast Cancer Cells ◽  
Monolayer Culture ◽  
Three Dimensional ◽  
Collagen Matrix ◽  
Living Cells ◽  
Two Dimensional ◽  
Metabolic Activities

The majority of in vitro studies of living cells are routinely conducted in a two-dimensional (2D) monolayer culture. Recent studies, however, suggest that 2D cell culture promotes specific types of...

scholarly journals Measurement and analysis of sarcomere length in rat cardiomyocytes in situ and in vitro

2010 ◽  
Vol 298 (5) ◽  
pp. H1616-H1625 ◽  
Author(s):  
G. Bub ◽  
P. Camelliti ◽  
C. Bollensdorff ◽  
D. J. Stuckey ◽  
G. Picton ◽  
...  
Keyword(s):  
Measurement Errors ◽  
Sarcomere Length ◽  
Ex Vivo ◽  
Living Cells ◽  
Perfused Heart ◽  
Experimental Conditions ◽  
Rat Cardiomyocytes ◽  
Two Photon

Sarcomere length (SL) is an important determinant and indicator of cardiac mechanical function; however, techniques for measuring SL in living, intact tissue are limited. Here, we present a technique that uses two-photon microscopy to directly image striations of living cells in cardioplegic conditions, both in situ (Langendorff-perfused rat hearts and ventricular tissue slices, stained with the fluorescent marker di-4-ANEPPS) and in vitro (acutely isolated rat ventricular myocytes). Software was developed to extract SL from two-photon fluorescence image sets while accounting for measurement errors associated with motion artifact in raster-scanned images and uncertainty of the cell angle relative to the imaging plane. Monte-Carlo simulations were used to guide analysis of SL measurements by determining error bounds as a function of measurement path length. The mode of the distribution of SL measurements in resting Langendorff-perfused heart is 1.95 μm ( n = 167 measurements from N = 11 hearts) after correction for tissue orientation, which was significantly greater than that in isolated cells (1.71 μm, n = 346, N = 9 isolations) or ventricular slice preparations (1.79 μm, n = 79, N = 3 hearts) under our experimental conditions. Furthermore, we find that edema in arrested Langendorff-perfused heart is associated with a mean SL increase; this occurs as a function of time ex vivo and correlates with tissue volume changes determined by magnetic resonance imaging. Our results highlight that the proposed method can be used to monitor SL in living cells and that different experimental models from the same species may display significantly different SL values under otherwise comparable conditions, which has implications for experiment design, as well as comparison and interpretation of data.

scholarly journals Three-dimensional migration of macrophages requires Hck for podosome organization and extracellular matrix proteolysis

Blood ◽  
2010 ◽  
Vol 115 (7) ◽  
pp. 1444-1452 ◽  
Author(s):  
Céline Cougoule ◽  
Véronique Le Cabec ◽  
Renaud Poincloux ◽  
Talal Al Saati ◽  
Jean-Louis Mège ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Spatial Organization ◽  
Ectopic Expression ◽  
Three Dimensional ◽  
Matrix Metalloproteases ◽  
3 Dimensional ◽  
Normal Expression ◽  
Migration Of Macrophages

Abstract Tissue infiltration of phagocytes exacerbates several human pathologies including chronic inflammations or cancers. However, the mechanisms involved in macrophage migration through interstitial tissues are poorly understood. We investigated the role of Hck, a Src-family kinase involved in the organization of matrix adhesion and degradation structures called podosomes. In Hck−/− mice submitted to peritonitis, we found that macrophages accumulated in interstitial tissues and barely reached the peritoneal cavity. In vitro, 3-dimensional (3D) migration and matrix degradation abilities, 2 protease-dependent properties of bone marrow–derived macrophages (BMDMs), were affected in Hck−/− BMDMs. These macrophages formed few and undersized podosome rosettes and, consequently, had reduced matrix proteolysis operating underneath despite normal expression and activity of matrix metalloproteases. Finally, in fibroblasts unable to infiltrate matrix, ectopic expression of Hck provided the gain–of–3D migration function, which correlated positively with formation of podosome rosettes. In conclusion, spatial organization of podosomes as large rosettes, proteolytic degradation of extracellular matrix, and 3D migration appeared to be functionally linked and regulated by Hck in macrophages. Hck, as the first protein combining a phagocyte-limited expression with a role in 3D migration, could be a target for new anti-inflammatory and antitumor molecules.

Nanofiber/hydrogel Core-shell Scaffolds With Three-dimensional Multilayer Patterned Structure for Accelerating Diabetic Wound Healing

2021 ◽  
Author(s):  
Jiankai Li ◽  
Tianshuai Zhang ◽  
Mingmang Pan ◽  
Feng Xue ◽  
Fang Lv ◽  
...  
Keyword(s):  
Wound Healing ◽  
Three Dimensional ◽  
Vapor Permeability ◽  
Core Shell ◽  
Diabetic Wound ◽  
3D Network ◽  
Diabetic Wound Healing ◽  
Diabetic Wounds

Abstract Impaired angiogenesis is one of the predominant reasons for non-healing diabetic wounds. Herein, a nanofiber/ hydrogel core-shell scaffold with three-dimensional (3D) multilayer patterned structure (3D-PT-P/GM) was introduced for promoting diabetic wound healing with improved angiogenesis. The results showed that the 3D-PT-P/GM scaffolds possessed multilayered structure with interlayer spacing of about 15-80 μm, and the hexagonal micropatterned structures were uniformly distributed on the surface of each layer. The nanofibers in the scaffold exhibited distinct core-shell structures with Gelatin methacryloyl (GelMA) hydrogel as the shell and Poly (D, L-lactic acid) (PDLLA) as the core. The results showed that the porosity, water retention time and water vapor permeability of the 3D-PT-P/GM scaffolds increased to 1.6 times, 21 times, and 1.9 times than that of the two-dimensional (2D) PDLLA nanofibrous scaffolds, respectively. The in vitro studies showed that the 3D-PT-P/GM scaffolds could significantly promote cell adhesion, proliferation, infiltration and migration throughout the scaffolds, and the expression of cellular communication protein-related genes, as well as angiogenesis-related genes in the same group, was remarkably upregulated. The in vivo results further demonstrated that the 3D-PT-P/GM scaffolds could not only effectively absorb exudate and provide a moist environment for the wound sites, but also significantly promote the formation of a 3D network of capillaries. As a result, the healing of diabetic wounds was accelerated with enhanced angiogenesis, granulation tissue formation, and collagen deposition. These results indicate that nanofiber/ hydrogel core-shell scaffolds with 3D multilayer patterned structures could provide a new strategy for facilitating chronic wound healing.

scholarly journals Structure of hibernating ribosomes studied by cryoelectron tomography in vitro and in situ

2010 ◽  
Vol 190 (4) ◽  
pp. 613-621 ◽  
Author(s):  
Julio O. Ortiz ◽  
Florian Brandt ◽  
Valério R.F. Matias ◽  
Lau Sennels ◽  
Juri Rappsilber ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Stress Response ◽  
Spatial Organization ◽  
Three Dimensional ◽  
E Coli ◽  
Escherichia Coli Cells ◽  
Three Dimensional Configuration

Ribosomes arranged in pairs (100S) have been related with nutritional stress response and are believed to represent a “hibernation state.” Several proteins have been identified that are associated with 100S ribosomes but their spatial organization has hitherto not been characterized. We have used cryoelectron tomography to reveal the three-dimensional configuration of 100S ribosomes isolated from starved Escherichia coli cells and we have described their mode of interaction. In situ studies with intact E. coli cells allowed us to demonstrate that 100S ribosomes do exist in vivo and represent an easily reversible state of quiescence; they readily vanish when the growth medium is replenished.

Laser-based 3D cell printing for tissue engineering

2014 ◽  
Vol 15 (3-4) ◽  
Author(s):  
Lothar Koch ◽  
Andrea Deiwick ◽  
Boris Chichkov
Keyword(s):  
Tissue Engineering ◽  
3D Printing ◽  
Three Dimensional ◽  
Living Cells ◽  
Cell Printing ◽  
Printing Technique ◽  
Cell Patterns ◽  
Printing Techniques

AbstractCurrently, different 3D printing techniques are investigated for printing biomaterials and living cells. An ambitious aim is the printing of fully functional tissue or organs. Furthermore, for manifold applications in biomedical research and in testing of pharmaceuticals or cosmetics, printed tissue could be a new method, partly substituting test animals. Here we describe a laser-based printing technique applied for the arrangement of vital cells in two and three-dimensional patterns and for tissue engineering. First printed tissue, tested in vitro and in vivo, and printing of cell patterns for investigating cell-cell interactions are presented.

scholarly journals Human iPS-derived pre-epicardial cells direct cardiomyocyte aggregation expansion and organization in vitro

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jun Jie Tan ◽  
Jacques P. Guyette ◽  
Kenji Miki ◽  
Ling Xiao ◽  
Gurbani Kaur ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Spatial Organization ◽  
Three Dimensional ◽  
Calcium Handling ◽  
Lateral Plate Mesoderm ◽  
Lateral Plate ◽  
Epicardial Cells ◽  
Tissue Organization ◽  
Cell Layers

AbstractEpicardial formation is necessary for normal myocardial morphogenesis. Here, we show that differentiating hiPSC-derived lateral plate mesoderm with BMP4, RA and VEGF (BVR) can generate a premature form of epicardial cells (termed pre-epicardial cells, PECs) expressing WT1, TBX18, SEMA3D, and SCX within 7 days. BVR stimulation after Wnt inhibition of LPM demonstrates co-differentiation and spatial organization of PECs and cardiomyocytes (CMs) in a single 2D culture. Co-culture consolidates CMs into dense aggregates, which then form a connected beating syncytium with enhanced contractility and calcium handling; while PECs become more mature with significant upregulation of UPK1B, ITGA4, and ALDH1A2 expressions. Our study also demonstrates that PECs secrete IGF2 and stimulate CM proliferation in co-culture. Three-dimensional PEC-CM spheroid co-cultures form outer smooth muscle cell layers on cardiac micro-tissues with organized internal luminal structures. These characteristics suggest PECs could play a key role in enhancing tissue organization within engineered cardiac constructs in vitro.

scholarly journals Bioprinting on 3D Printed Titanium Scaffolds for Periodontal Ligament Regeneration

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1337
Author(s):  
Ui-Lyong Lee ◽  
Seokhwan Yun ◽  
Hua-Lian Cao ◽  
Geunseon Ahn ◽  
Jin-Hyung Shim ◽  
...  
Keyword(s):  
Dental Implants ◽  
Periodontal Ligament ◽  
Defense Mechanisms ◽  
Animal Experiments ◽  
Three Dimensional ◽  
Artificial Organs ◽  
Artificial Organ ◽  
Mechanical Stimuli ◽  
3D Printed

The three-dimensional (3D) cell-printing technique has been identified as a new biofabrication platform because of its ability to locate living cells in pre-defined spatial locations with scaffolds and various growth factors. Osseointegrated dental implants have been regarded as very reliable and have long-term reliability. However, host defense mechanisms against infections and micro-movements have been known to be impaired around a dental implant because of the lack of a periodontal ligament. In this study, we fabricated a hybrid artificial organ with a periodontal ligament on the surface of titanium using 3D printing technology. CEMP-1, a known cementogenic factor, was enhanced in vitro. In animal experiments, when the hybrid artificial organ was transplanted to the calvarial defect model, it was observed that the amount of connective tissue increased. 3D-printed hybrid artificial organs can be used with dental implants, establishing physiological tooth functions, including the ability to react to mechanical stimuli and the ability to resist infections.

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