A Device for Long Term, In Vitro Loading of Three-Dimensional Natural and Engineered Tissues

2003 ◽  
Vol 31 (11) ◽  
pp. 1347-1356 ◽  
Author(s):  
Daniel A. Shimko ◽  
Kyle K. White ◽  
Eric A. Nauman ◽  
Kay C Dee
Keyword(s):  
Three Dimensional ◽  
Engineered Tissues

Long-term in vitro human pancreatic islet culture using three-dimensional microfabricated scaffolds

Biomaterials ◽  
2011 ◽  
Vol 32 (6) ◽  
pp. 1536-1542 ◽  
Author(s):  
Jamal T. Daoud ◽  
Maria S. Petropavlovskaia ◽  
Jason M. Patapas ◽  
Christian E. Degrandpré ◽  
Robert W. DiRaddo ◽  
...  
Keyword(s):  
Pancreatic Islet ◽  
Three Dimensional ◽  
Human Pancreatic Islet ◽  
Islet Culture

scholarly journals Altered swelling and ion fluxes in articular cartilage as a biomarker in osteoarthritis and joint immobilization: a computational analysis

2015 ◽  
Vol 12 (102) ◽  
pp. 20141090 ◽  
Author(s):  
Sara Manzano ◽  
Raquel Manzano ◽  
Manuel Doblaré ◽  
Mohamed Hamdy Doweidar
Keyword(s):  
Articular Cartilage ◽  
Computational Model ◽  
Three Dimensional ◽  
Ion Distribution ◽  
Tissue Degeneration ◽  
Structural Deterioration ◽  
Healthy Cartilage ◽  
Tissue Degradation

In healthy cartilage, mechano-electrochemical phenomena act together to maintain tissue homeostasis. Osteoarthritis (OA) and degenerative diseases disrupt this biological equilibrium by causing structural deterioration and subsequent dysfunction of the tissue. Swelling and ion flux alteration as well as abnormal ion distribution are proposed as primary indicators of tissue degradation. In this paper, we present an extension of a previous three-dimensional computational model of the cartilage behaviour developed by the authors to simulate the contribution of the main tissue components in its behaviour. The model considers the mechano-electrochemical events as concurrent phenomena in a three-dimensional environment. This model has been extended here to include the effect of repulsion of negative charges attached to proteoglycans. Moreover, we have studied the fluctuation of these charges owning to proteoglycan variations in healthy and pathological articular cartilage. In this sense, standard patterns of healthy and degraded tissue behaviour can be obtained which could be a helpful diagnostic tool. By introducing measured properties of unhealthy cartilage into the computational model, the severity of tissue degeneration can be predicted avoiding complex tissue extraction and subsequent in vitro analysis. In this work, the model has been applied to monitor and analyse cartilage behaviour at different stages of OA and in both short (four, six and eight weeks) and long-term (11 weeks) fully immobilized joints. Simulation results showed marked differences in the corresponding swelling phenomena, in outgoing cation fluxes and in cation distributions. Furthermore, long-term immobilized patients display similar swelling as well as fluxes and distribution of cations to patients in the early stages of OA, thus, preventive treatments are highly recommended to avoid tissue deterioration.

scholarly journals The Bordetella Bps Polysaccharide Is Critical for Biofilm Development in the Mouse Respiratory Tract

2007 ◽  
Vol 189 (22) ◽  
pp. 8270-8276 ◽  
Author(s):  
Gina Parise Sloan ◽  
Cheraton F. Love ◽  
Neelima Sukumar ◽  
Meenu Mishra ◽  
Rajendar Deora
Keyword(s):  
Whooping Cough ◽  
Three Dimensional ◽  
Respiratory Pathogens ◽  
Adolescents And Adults ◽  
The Stability ◽  
Inert Surfaces ◽  
Biofilm Development

ABSTRACT Bordetellae are respiratory pathogens that infect both humans and animals. Bordetella bronchiseptica establishes asymptomatic and long-term to life-long infections of animal nasopharynges. While the human pathogen Bordetella pertussis is the etiological agent of the acute disease whooping cough in infants and young children, it is now being increasingly isolated from the nasopharynges of vaccinated adolescents and adults who sometimes show milder symptoms, such as prolonged cough illness. Although it has been shown that Bordetella can form biofilms in vitro, nothing is known about its biofilm mode of existence in mammalian hosts. Using indirect immunofluorescence and scanning electron microscopy, we examined nasal tissues from mice infected with B. bronchiseptica. Our results demonstrate that a wild-type strain formed robust biofilms that were adherent to the nasal epithelium and displayed architectural attributes characteristic of a number of bacterial biofilms formed on inert surfaces. We have previously shown that the Bordetella Bps polysaccharide encoded by the bpsABCD locus is critical for the stability and maintenance of three-dimensional structures of biofilms. We show here that Bps is essential for the formation of efficient nasal biofilms and is required for the colonization of the nose. Our results document a biofilm lifestyle for Bordetella in mammalian respiratory tracts and highlight the essential role of the Bps polysaccharide in this process and in persistence of the nares.

scholarly journals Investigating PEGDA and GelMA Microgel Models for Sustained 3D Heterotypic Dermal Papilla and Keratinocyte Co-Cultures

2021 ◽  
Vol 22 (4) ◽  
pp. 2143 ◽  
Author(s):  
Justin J.Y. Tan ◽  
Duc-Viet Nguyen ◽  
John E. Common ◽  
Chunyong Wu ◽  
Paul C.L. Ho ◽  
...  
Keyword(s):  
Hair Follicle ◽  
Three Dimensional ◽  
Dermal Papilla ◽  
Glycol Diacrylate ◽  
Prolonged Duration ◽  
Culture Models ◽  
Poly Ethylene

Hair follicle morphogenesis is heavily dependent on reciprocal, sequential, and epithelial-mesenchymal interaction (EMI) between epidermal stem cells and the specialized cells of the underlying mesenchyme, which aggregate to form the dermal condensate (DC) and will later become the dermal papilla (DP). Similar models were developed with a co-culture of keratinocytes and DP cells. Previous studies have demonstrated that co-culture with keratinocytes maintains the in vivo characteristics of the DP. However, it is often challenging to develop three-dimensional (3D) DP and keratinocyte co-culture models for long term in vitro studies, due to the poor intercellular adherence between keratinocytes. Keratinocytes exhibit exfoliative behavior, and the integrity of the DP and keratinocyte co-cultured spheroids cannot be maintained over prolonged culture. Short durations of culture are unable to sufficiently allow the differentiation and re-programming of the keratinocytes into hair follicular fate by the DP. In this study, we explored a microgel array approach fabricated with two different hydrogel systems. Using poly (ethylene glycol) diacrylate (PEGDA) and gelatin methacrylate (GelMA), we compare their effects on maintaining the integrity of the cultures and their expression of important genes responsible for hair follicle morphogenesis, namely Wnt10A, Wnt10B, and Shh, over prolonged duration. We discovered that low attachment surfaces such as PEGDA result in the exfoliation of keratinocytes and were not suitable for long-term culture. GelMA, on the hand, was able to sustain the integrity of co-cultures and showed higher expression of the morphogens overtime.

scholarly journals Long-term maintenance of functional primary human hepatocytes in 3D gelatin matrices produced by solution blow spinning

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mariliis Klaas ◽  
Kaidi Möll ◽  
Kristina Mäemets-Allas ◽  
Mart Loog ◽  
Martin Järvekülg ◽  
...  
Keyword(s):  
Liver Tissue ◽  
Drug Testing ◽  
Three Dimensional ◽  
Human Hepatocytes ◽  
Drug Induced ◽  
Human Liver Tissue ◽  
Drug Induced Liver Injury ◽  
Solution Blow Spinning

AbstractSolution blow spinning (SBS) has recently emerged as a novel method that can produce nano- and microfiber structures suitable for tissue engineering. Gelatin is an excellent precursor for SBS as it is derived mainly from collagens that are abundant in natural extracellular matrices. Here we report, for the first time the successful generation of 3D thermally crosslinked preforms by using SBS from porcine gelatin. These SBS mats were shown to have three-dimensional fibrous porous structure similar to that of mammalian tissue extracellular matrix. In pharma industry, there is an urgent need for adequate 3D liver tissue models that could be used in high throughput setting for drug screening and to assess drug induced liver injury. We used SBS mats as culturing substrates for human hepatocytes to create an array of 3D human liver tissue equivalents in 96-well format. The SBS mats were highly cytocompatible, facilitated the induction of hepatocyte specific CYP gene expression in response to common medications, and supported the maintenance of hepatocyte differentiation and polarization status in long term cultures for more than 3 weeks. Together, our results show that SBS-generated gelatin scaffolds are a simple and efficient platform for use in vitro for drug testing applications.

scholarly journals Microfabricated disk technology: rapid scale up in midbrain organoid generation

2021 ◽  
Author(s):  
Nguyen-Vi Mohamed ◽  
Paula Lepine ◽  
Maria Lacalle-Aurioles ◽  
Julien Sirois ◽  
Meghna Mathur ◽  
...  
Keyword(s):  
Scale Up ◽  
Three Dimensional ◽  
Require Time ◽  
Large Numbers ◽  
Midbrain Region ◽  
Brain Organoid ◽  
Induced Pluripotent ◽  
The Brain

By providing a three-dimensional in vitro culture system with key features of the substantia nigra region in the brain, 3D neuronal organoids derived from human induced pluripotent stem cells (iPSCs) provide living neuronal tissue resembling the midbrain region of the brain. However, a major limitation of conventional brain organoid culture is that it is often labor-intensive, requiring highly specialized personnel for moderate throughput. Additionally, the methods published for long-term cultures require time-consuming maintenance to generate brain organoids in large numbers. With the increasing need for human midbrain organoids (hMOs) to better understand and model Parkinson′s disease (PD) in a dish, there is a need to implement new workflows and methods to both generate and maintain hMOs, while minimizing batch to batch variation. In this study, we developed a method with microfabricated disks to scale up the generation of hMOs. This opens up the possibility to generate larger numbers of hMOs, in a manner that minimizes the amount of labor required, while decreasing variability and maintaining the viability of these hMOs over time. Taken together, producing hMOs in this manner opens up the potential for these to be used to further PD studies.

Three-dimensional Co-culture of Primary Human Liver Cells in Bioreactors for In Vitro Drug Studies: Effects of the Initial Cell Quality on the Long-term Maintenance of Hepatocyte-specific Functions

2002 ◽  
Vol 30 (5) ◽  
pp. 525-538 ◽  
Author(s):  
Katrin Zeilinger ◽  
Igor M. Sauer ◽  
Gesine Pless ◽  
Catrin Strobel ◽  
Jeannette Rudzitis ◽  
...  
Keyword(s):  
Pharmaceutical Industry ◽  
Human Liver ◽  
Three Dimensional ◽  
Liver Cells ◽  
Bioreactor Culture ◽  
Initial Cell ◽  
Human Liver Cells ◽  
Term Maintenance

In vitro culture models that employ human liver cells could be potent tools for predictive studies on drug toxicity and metabolism in the pharmaceutical industry. A bioreactor culture model was developed that permits the three-dimensional co-culture of liver cells under continuous medium perfusion with decentralised mass exchange and integral oxygenation. We tested the ability of the system to support the long-term maintenance and differentiation of primary human liver cells. The effects of the initial cell quality were investigated by comparing cultures from resected, non-preserved liver with cultures from liver graft tissue damaged by long-term preservation. In cultures originating from non-preserved liver, protein and urea synthesis, glucose metabolism, and cytochrome (P450) activities were stable over the 2-week culture period, with maximal activities at the end of the first week in culture. Enzyme induction led to increased 7-ethoxyresorufin O-deethylase activities of up to 20 times the basal value. In cultures from preservation-damaged liver, recovery of metabolic activities was detected during bioreactor culture. After two weeks, most biochemical parameters approached those of cultures from non-preserved human liver. Light microscopy demonstrated the three-dimensional reorganisation of hepatocytes and nonparenchymal cells in co-culture. Long-term maintenance, and even the regeneration of specific functional activities of human liver cells, can be achieved in the bioreactor. This could facilitate the introduction into the pharmaceutical industry of in vitro drug testing with primary human liver cells.

scholarly journals Novel Bioengineered Three-Dimensional Human Intestinal Model for Long-Term Infection of Cryptosporidium parvum

2017 ◽  
Vol 85 (3) ◽  
Author(s):  
Maria A DeCicco RePass ◽  
Ying Chen ◽  
Yinan Lin ◽  
Wenda Zhou ◽  
David L. Kaplan ◽  
...  
Keyword(s):  
Cryptosporidium Parvum ◽  
Drug Targets ◽  
Diarrheal Disease ◽  
Three Dimensional ◽  
Content Type ◽  
Tissue Model ◽  
Culture Models

ABSTRACT Cryptosporidium spp. are apicomplexan parasites of global importance that cause human diarrheal disease. In vitro culture models that may be used to study this parasite and that have physiological relevance to in vivo infection remain suboptimal. Thus, the pathogenesis of cryptosporidiosis remains poorly characterized, and interventions for the disease are limited. In this study, we evaluated the potential of a novel bioengineered three-dimensional (3D) human intestinal tissue model (which we developed previously) to support long-term infection by Cryptosporidium parvum. Infection was assessed by immunofluorescence assays and confocal and scanning electron microscopy and quantified by quantitative reverse transcription-PCR. We found that C. parvum infected and developed in this tissue model for at least 17 days, the extent of the study time used in the present study. Contents from infected scaffolds could be transferred to fresh scaffolds to establish new infections for at least three rounds. Asexual and sexual stages and the formation of new oocysts were observed during the course of infection. Additionally, we observed ablation, blunting, or distortion of microvilli in infected epithelial cells. Ultimately, a 3D model system capable of supporting continuous Cryptosporidium infection will be a useful tool for the study of host-parasite interactions, identification of putative drug targets, screening of potential interventions, and propagation of genetically modified parasites.

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