Proliferation and Interactions of Several Cell Types Encapsulated Within Dense (Non-Porous) Protein-Permeable Polyurethane Membranes

1993 ◽  
Vol 331 ◽  
Author(s):  
Albert Y. Wang ◽  
Robert S. Ward ◽  
Kathleen A. White ◽  
Robert W. Kuhn ◽  
Julie E. Taylor ◽  
...  
Keyword(s):  
Fibrous Tissue ◽  
Cell Encapsulation ◽  
Cell Types ◽  
Artificial Organs ◽  
Encapsulated Cells ◽  
Polyurethane Membranes ◽  
Dependent Cell ◽  
And Function

AbstractProtein-permeable dense (non-porous) urethane membranes have been evaluated for in vitro cell culture, and in vivo cell encapsulation. Polyurethane membranes were designed to exhibit permeability to proteins, gases, and nutrients without the existence of pores. The membranes are non-cytotoxic, angiogenic, and permeable to gases, nutrients, secretagogues and cell products via purely concentration-driven transport. Non-anchorage and anchorage dependent cells were grown encapsulated within the membrane and with the membrane as a growth substrate. Several non-anchorage dependent cell types proliferated within the membrane both in-vitro and in-vivo. Anchorage-dependent cells were grown on the membranes as a substrate. Encapsulated cells have been maintained in culture for up to six months with nutrients supplied only by the external media. Immuno-isolation has been demonstrated with cells implanted into murine hosts. Explants of membrane encapsulated cells exhibited a high degree of vascularization, with little or no fibrous tissue. The ability to support cell growth and function, and the ability to protect xenogenic cells from immunologic rejection suggest that the membranes would be useful in the construction of hybrid artificial organs, devices for cell transplantation, and substrates for cell and tissue culture.

scholarly journals Modelling the Human Placental Interface In Vitro—A Review

Micromachines ◽  
2021 ◽  
Vol 12 (8) ◽  
pp. 884
Author(s):  
Marta Cherubini ◽  
Scott Erickson ◽  
Kristina Haase
Keyword(s):  
Drug Testing ◽  
Cell Types ◽  
In Vitro Models ◽  
Placental Development ◽  
Scientific Challenge ◽  
Induced Pluripotent ◽  
And Function ◽  
And Control

Acting as the primary link between mother and fetus, the placenta is involved in regulating nutrient, oxygen, and waste exchange; thus, healthy placental development is crucial for a successful pregnancy. In line with the increasing demands of the fetus, the placenta evolves throughout pregnancy, making it a particularly difficult organ to study. Research into placental development and dysfunction poses a unique scientific challenge due to ethical constraints and the differences in morphology and function that exist between species. Recently, there have been increased efforts towards generating in vitro models of the human placenta. Advancements in the differentiation of human induced pluripotent stem cells (hiPSCs), microfluidics, and bioprinting have each contributed to the development of new models, which can be designed to closely match physiological in vivo conditions. By including relevant placental cell types and control over the microenvironment, these new in vitro models promise to reveal clues to the pathogenesis of placental dysfunction and facilitate drug testing across the maternal–fetal interface. In this minireview, we aim to highlight current in vitro placental models and their applications in the study of disease and discuss future avenues for these in vitro models.

scholarly journals Polymorphonuclear Leukocytes Prepared by Continuous-Flow Filtration Leukapheresis: Viability and Function

Blood ◽  
1974 ◽  
Vol 44 (5) ◽  
pp. 707-713 ◽  
Author(s):  
Michael B. Harris ◽  
Isaac Djerassi ◽  
Elias Schwartz ◽  
Richard K. Root
Keyword(s):  
Continuous Flow ◽  
Polymorphonuclear Leukocytes ◽  
Cell Types ◽  
High Yield ◽  
Trypan Blue Exclusion ◽  
Flow Filtration ◽  
And Function ◽  
Combating Infection

Abstract Preparation of granulocytes for transfusion in high yield and relatively free of contamination by other cell types has been made possible by the technique of continuous-flow filtration leukapheresis (CFFL). Since previous work suggested that granulocytes collected in this manner may have impaired viability and function, a detailed study of the bactericidal, metabolic, and chemotactic properties of such cells was performed and compared to control cells obtained from the same donors prior to CFFL. The granulocyte percentage of the cell suspensions obtained by CFFL averaged 94.5% ± 1.5% compared to 82.5% ± 1.8% for the controls (p < 0.001) with viability of the PMNs determined by trypan blue exclusion being 97.5% ± 0.9% and 98.2% ± 0.5%, respectively. The phogocytic, metabolic (14C-I-glucose oxidation and protein iodination) and chemotactic properties of both cell types were equivalent in suspensions equalized for granulocyte content. These findings indicate that CFFL technique employed does not impair granulocyte viability or function in vitro. Studies of the in vivo survival and function of CFFL granulocytes are necessary to evaluate their efficacy in combating infection in severely leukopenic patients.

P0658MATRIX AND FLOW MODULATE GENE EXPRESSION IN A 3D VASCULARISED TUBULE MODEL

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Julie Williams ◽  
Sanlin Robinson ◽  
Babak Alaei ◽  
Kimberly Homan ◽  
Maryam Clausen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Impact Analysis ◽  
Cell Types ◽  
Drug Uptake ◽  
Shear Stresses ◽  
The Matrix ◽  
And Function

Abstract Background and Aims Questions abound regarding the translation of in vitro 2D cell culture systems to the human setting. This is especially true of the kidney in which there is a complex hierarchical structure and a multitude of cell types. While it is well accepted that extracellular matrix plays a large part in directing cellular physiology emerging research has highlighted the importance of shear stresses and flow rates too. To fully recapitulate the normal gene expression and function of a particular renal cell type how important is it to completely reconstitute their in vivo surroundings? Method To answer this question, we have cultured proximal tubular (PT) epithelial cells in a 3-dimensional channel embedded within an engineered extracellular matrix (ECM) under physiological flow that is colocalised with an adjacent channel lined with renal microvascular endothelial cells that mimic a peritubular capillary. Modifications to the system were made to allow up to 12 chips to be run in parallel in an easily handleable form. After a period of maturation under continuous flow, both cell types were harvested for RNAseq analyses. RNA expression data was compared with cells cultured under static 2-dimensional conditions on plastic or the engineered ECM. Additionally, the perfusion of glucose through this 3D vascularised PT model has been investigated in the presence and absence of known diabetes modulating agents. Results PCA of RNAseq data showed that a) static non-coated, b) static matrix-coated and c) flow matrix-coated conditions separated into 3 distinct groups, while cell co-culture had less impact. Analysis of transcriptomic signatures showed that many genes were modulated by the matrix with additional genes influenced under flow conditions. Several of these genes, classified as transporters, are of particular importance when using this model to assess drug uptake and safety implications. Co-culture regulated some interesting genes, but fewer than anticipated. Preliminary experiments are underway to monitor glucose uptake and transport between tubules under different conditions. Conclusion We have developed a medium throughput system in which matrix and flow modulate gene expression. This system can be used to study the physiology of molecular cross-talk between cells. Ongoing analysis will further consider relevance to human physiology.

scholarly journals Antibody-dependent eosinophil-mediated damage to 51Cr-labeled schistosomula of Schistosoma mansoni: damage by purieid eosinophils.

1977 ◽  
Vol 145 (1) ◽  
pp. 136-150 ◽  
Author(s):  
A E Butterworth ◽  
J R David ◽  
D Franks ◽  
A A Mahmoud ◽  
P H David ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Blood Leukocytes ◽  
Normal Individuals ◽  
Dependent Cell ◽  
A Cell ◽  
Relationship Of

After earlier observations that antibody-dependent, cell-mediated damage to 51Cr-labeled schistosomula can be ablated by pretreatment of a mixed preparation of human peripheral blood leukocytes with an anti-eosinophil serum and complement, we investigated the cytotoxic effects of eosinophil-enriched cell preparations. Preparations containing up to 98.5% eosinophils and devoid of neutrophils were effective in mediating antibody-dependent damage to schistosomula. Preparations enriched in mononuclear cells or in neutrophils, and devoid of eosinophils, were inactive. Eosinophils from some patients with eosinophilia induced by schistosomiasis were less active on a cell-to-cell basis than cells from normal individuals. The possibility that such cells were initially blocked by immune complexes was considered, and it was found that reasonable cytotoxicity by purified eosinophils from patients with eosinophilia could be generated by overnight cultures. A possible requirement for cooperation between eosinophils and other cell types was also studied. Lymphocytes, neutrophils and monocytes failed to enhance eosinophil-mediated cytotoxicity. These results provide further evidence that the eosinophil is the only cell in man responsible for antibody-dependent, complement-independent damage to schistosomula in vitro. Eosinophils from individuals, however, differ in their cytotoxic potential by a mechanism yet to be elucidated. The possible relationship of these findings to immunity in vivo is discussed.

scholarly journals Single-cell transcriptomics reveals multiple neuronal cell types in human midbrain-specific organoids

2019 ◽  
Author(s):  
Lisa M. Smits ◽  
Stefano Magni ◽  
Kamil Grzyb ◽  
Paul MA. Antony ◽  
Rejko Krüger ◽  
...  
Keyword(s):  
Single Cell ◽  
Dopaminergic Neurons ◽  
Neuronal Cell ◽  
Cell Types ◽  
Human Stem Cell ◽  
Glutamatergic Neurons ◽  
And Function ◽  
Excellent Tool

AbstractHuman stem cell-derived organoids have great potential for modelling physiological and pathological processes. They recapitulatein vitrothe organisation and function of a respective organ or part of an organ. Human midbrain organoids (hMOs) have been described to contain midbrain-specific dopaminergic neurons that release the neurotransmitter dopamine. However, the human midbrain contains also additional neuronal cell types, which are functionally interacting with each other. Here, we analysed hMOs at high-resolution by means of single-cell RNA-sequencing (scRNA-seq), imaging and electrophysiology to unravel cell heterogeneity. Our findings demonstrate that hMOs show essential neuronal functional properties as spontaneous electrophysiological activity of different neuronal subtypes, including dopaminergic, GABAergic, and glutamatergic neurons. Recapitulating thesein vivofeatures makes hMOs an excellent tool forin vitrodisease phenotyping and drug discovery.

scholarly journals Signaling roles of phosphoinositides in the retina

2020 ◽  
pp. jlr.TR120000806 ◽  
Author(s):  
Raju V. S. Rajala
Keyword(s):  
Cell Types ◽  
Cellular Functions ◽  
Parent Molecule ◽  
Phosphoinositide Signaling ◽  
Wide Range ◽  
The Many ◽  
And Function ◽  
Different Cell Types

The field of phosphoinositide signaling has expanded significantly in recent years. Phosphoinositides (PIs) are universal signaling molecules that directly interact with membrane proteins or with cytosolic proteins containing domains that directly bind phosphoinositides and are recruited to cell membranes. Through the activities of PI kinases and PI phosphatases, seven distinct phosphoinositide lipid molecules are formed from the parent molecule phosphatidylinositol. PI signals regulate a wide range of cellular functions, including cytoskeletal assembly, membrane binding and fusion, ciliogenesis, vesicular transport, and signal transduction. Given the many excellent reviews on phosphoinositide kinases, phosphoinositide phosphatases, and PIs in general, in this review, we discuss recent studies and advances in PI lipid signaling in the retina. We specifically focus on PI lipids from vertebrate (e.g. bovine, rat, mice, toad, and zebrafish) and invertebrate (e.g. drosophila, horseshoe crab, and squid) retinas. We also discuss the importance of PIs revealed from animal models and human diseases, and methods to study PI levels both in vitro and in vivo. We propose that future studies should investigate the function and mechanism of activation of PI-modifying enzymes/phosphatases and further unravel PI regulation and function in the different cell types of the retina.

scholarly journals Cross-species Transcriptomic Comparison of In Vitro and In Vivo Mammalian Neural Cells

2015 ◽  
Vol 9 ◽  
pp. BBI.S33124 ◽  
Author(s):  
Peter R. LoVerso ◽  
Christopher M. Wachter ◽  
Feng Cui
Keyword(s):  
Gene Expression ◽  
Transcript Abundance ◽  
Cell Types ◽  
Mammalian Brain ◽  
Neural Cells ◽  
Rna Seq ◽  
Commercial Sources ◽  
And Function

The mammalian brain is characterized by distinct classes of cells that differ in morphology, structure, signaling, and function. Dysregulation of gene expression in these cell populations leads to various neurological disorders. Neural cells often need to be acutely purified from animal brains for research, which requires complicated procedure and specific expertise. Primary culture of these cells in vitro is a viable alternative, but the differences in gene expression of cells grown in vitro and in vivo remain unclear. Here, we cultured three major neural cell classes of rat brain (ie, neurons, astrocytes, and oligodendrocyte precursor cells [OPCs]) obtained from commercial sources. We measured transcript abundance of these cell types by RNA sequencing (RNA-seq) and compared with their counterparts acutely purified from mouse brains. Cross-species RNA-seq data analysis revealed hundreds of genes that are differentially expressed between the cultured and acutely purified cells. Astrocytes have more such genes compared to neurons and OPCs, indicating that signaling pathways are greatly perturbed in cultured astrocytes. This dataset provides a powerful resource to demonstrate the similarities and differences of biological processes in mammalian neural cells grown in vitro and in vivo at the molecular level.

A Novel Suspended Hydrogel Membrane Platform for Cell Culture

2015 ◽  
Vol 6 (2) ◽  
Author(s):  
Yong X. Chen ◽  
Shihao Yang ◽  
Jiahan Yan ◽  
Ming-Han Hsieh ◽  
Lingyan Weng ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Biology ◽  
3D Cell Culture ◽  
Cell Encapsulation ◽  
Cell Types ◽  
Ease Of Use ◽  
In Vitro Assays ◽  
Hydrogel Membrane

Current cell-culture is largely performed on synthetic two-dimensional (2D) petri dishes or permeable supports such as Boyden chambers, mostly because of their ease of use and established protocols. It is generally accepted that modern cell biology research requires new physiologically relevant three-dimensional (3D) cell culture platform to mimic in vivo cell responses. To that end, we report the design and development of a suspended hydrogel membrane (ShyM) platform using gelatin methacrylate (GelMA) hydrogel. ShyM thickness (0.25–1 mm) and mechanical properties (10–70 kPa) can be varied by controlling the size of the supporting grid and concentration of GelMA prepolymer, respectively. GelMA ShyMs, with dual media exposure, were found to be compatible with both the cell-seeding and the cell-encapsulation approach as tested using murine 10T1/2 cells and demonstrated higher cellular spreading and proliferation as compared to flat GelMA unsuspended control. The utility of ShyM was also demonstrated using a case-study of invasion of cancer cells. ShyMs, similar to Boyden chambers, are compatible with standard well-plates designs and can be printed using commonly available 3D printers. In the future, ShyM can be potentially extended to variety of photosensitive hydrogels and cell types, to develop new in vitro assays to investigate complex cell–cell and cell–extracellular matrix (ECM) interactions.

scholarly journals Problems and prospects for the use of stem cells in transplantation

2021 ◽  
Vol 23 (2) ◽  
pp. 175-186
Author(s):  
Alexander V. Moskalev ◽  
Boris Yu. Gumilevsky ◽  
Vasiliy Ya. Apchel ◽  
Vasiliy N. Cygan
Keyword(s):  
Stem Cells ◽  
Cell Types ◽  
Artificial Organs ◽  
Immune Rejection ◽  
Intercellular Interaction ◽  
Genetic Changes ◽  
Technical Problems ◽  
The Moment

The problems of organ and tissue transplantation are the lack of organs for transplantation and the rejection of transplants. Therefore, the issue of obtaining organs and tissues for transplantation with stem cells is being studied. Although this idea is promising, it is associated with many problems. To do this, you need to use several populations of cells on a substrate with a complex composition of nutrient environments: nutrients, growth factors, oxygen, regulatory factors. Intercellular interaction is provided by the factors they secrete, or it occurs directly with intercellular contact. This contributes to the fact that stem cells in test tubes can differentiate into other types of tissues and maintain their biological activity indefinitely, which they cannot in vivo. This approach of tissue engineering provides the possibility of obtaining whole organs for implantation. However, technical problems are associated with increased cell adhesion to plastic, the presence of a universal basis for cell nutrition, which can contain more than 100 components. There is a possibility of contamination, which can lead to serious errors in the experiment. Stem cells must have distinct mutational properties and the ability to restore telome cells. Prolonged use of the same nutrient medium can lead to genetic changes and significantly alter the physiological properties of cells. Cryopreservation can be an important aspect of the solution. The goal of tissue bioengineering is to create whole artificial organs, or at least areas of organized tissue that could be transplanted to patients. Currently, such operations are relatively simple for tissues such as artificial skin consisting of epidermal and fibroblast layers, or small cartilage implants obtained in vitro. Several cell types in stable shape are planned to be used in one environment. In this case, one type of cell can be replaced by another. This stability is provided by a variety of secreted factors by different types of cells that ensure their vitality. Decellularization removes all components involved in immune rejection of grafts, so this raises the prospect of creating an unlimited supply of organs for transplantation. However, acute reactions can develop associated with the participation of dendritic cells, macrophages, neutrophils, natural killers. Starting from the moment of transplantation, conditions for immune rejection are created, arising as a result of surgery with the development of acute inflammation. The intensity of immune reactions against the graft largely depends on the degree of non-conformity of alleles of the main complex of histocompany capacity of the donor and recipient. This match is studied using a variety of methods, including the use of antibodies or sequencing of deoxyribonucleic acid.

Hepatocyte Labeling with 99mTc-GSA: A Potential Non-Invasive Technique for Tracking Cell Transplantation

2012 ◽  
Vol 35 (6) ◽  
pp. 450-457 ◽  
Author(s):  
Manil Chouhan ◽  
Juliana Puppi ◽  
Estela Solanas ◽  
Ragai R. Mitry ◽  
Anil Dhawan ◽  
...  
Keyword(s):  
Rat Hepatocytes ◽  
Fold Increase ◽  
Cell Types ◽  
Human Hepatocytes ◽  
Hepatocyte Transplantation ◽  
Invasive Technique ◽  
Promising Alternative ◽  
And Function

Background: Hepatocyte transplantation is a promising alternative to orthotopic liver transplantation, however, the fate of transplanted hepatocytes is not well defined. 99mTc-galactosyl-serum albumin (99mTc-GSA) is a clinical scintigraphic agent which is specifically taken up by the hepatocyte asialoglycoprotein receptor (ASGPR). Aims: To investigate labeling of fresh and cryopreserved human hepatocytes and fresh rat hepatocytes in vitro using 99mTc-GSA Methods: Human and rat hepatocytes were isolated from liver tissue by collagenase perfusion. The ASGPR were characterized using immunohistochemistry and RT-PCR. Hepatocytes were incubated with 99mTc-GSA in suspension at 4°C and 37°C. Cell viability and function was determined using cell mitochondrial dehydrogenase (MTS) and sulphorhodamine B (SRB) assays. Results: Fresh and cryopreserved human hepatocytes expressed the ASGPR. Incubation of hepatocytes in suspension with 99mTc-GSA reduced the viability of hepatocytes, but this was similar to unlabeled control cells. Greater loss of viability was seen on incubation at 37°C compared to 4°C, but there was a significantly greater uptake of 99mTc-GSA at the physiological temperature (6.6 ± SE 0.6-fold increase, p<0.05) consistent with ASGPR-mediated endocytosis. MTS and SRB assays were not significantly affected by labeling with 99mTc-GSA in all three cell types. A mean of 18.5% of the radioactivity was released over 120 min when 99mTc-GSA - labeled hepatocytes were shaken in vitro at 37°C. Conclusions: Human and rat hepatocytes can be labeled with 99mTc-GSA, which may have potential application for in vivo imaging after hepatocyte transplantation.

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