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Table 1 Donor characteristics and processing data from 33 clinical islet preparations. Open in a separate window. Morphological analysis A 0. Figure 1. Figure 2. Cell number fraction by EM Cell composition of an islet preparation was determined by counting the different cell types on the electron micrographs. Islet purity in a preparation determined by three methods Volume fraction islets determined using cell number fraction, cell volume, and extracellular volume fraction Number fraction data was converted to a volume fraction basis in order to compare with data from other techniques and from the literature.

Results Determination of cell composition and number fraction by EM Different cell types within a preparation were definitively identified by their known ultrastructural characteristics Figure 2. Table 3 Number fraction of cell types in 33 islet preparations determined by EM. Cell volume fraction determination The cell number fraction data determined by EM Table 3 and estimates of the volume of each cell type were used with Equations 7 - 9 and 13 - 15 of the Appendix to obtain the average cell volume for islet and non-islet cells and for the total of all cells. Table 4 Volume fraction of cell types in 33 islet preparations determined by EM and the theoretical number of cells in an islet equivalent IE.

Islet volume fraction evaluated by consideration of extracellular volume It is important to make the distinction between the volume of cells in an islet and the total tissue volume of an islet; the latter includes extracellular volume, which results in a larger volume. Figure 3. Comparison of islet volume fractions determined by three methods Transplanted human islet preparations are typically accompanied by many acinar and duct cells.

Figure 4. Figure 5. Figure 6. Figure 7. Discussion The islet transplantation field is challenged with the need of finding better ways to standardize methods for determining the composition of islet preparations. All other studies with pancreatic blocks did not include PP-rich lobes.

Appendix Number fraction and volume fraction relationships in islet preparation Our objective is to develop a framework of equations and parameters for conversion between number fraction and volume fraction measurements with human pancreatic islet preparations.

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Cells Cell composition measurements from EM provide data on the number n i of each type of cell in the sample, where i represents the cell type. Tissues Next we develop expressions for volume fractions based upon the total tissue volumes, V I and V NI , the components of which are shown schematically in Figure 9. Figure 8. Parameters Estimates of the cell volumes and tissue extracellular volumes are needed to carry out the calculations described here. References 1.


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Single-donor, marginal-dose islet transplantation in patients with type 1 diabetes. Five-year follow-up after clinical islet transplantation. Islet graft assessment in the Edmonton Protocol: implications for predicting long-term clinical outcome.

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Refinement of the automated method for human islet isolation and presentation of a closed system for in vitro islet culture. Implantation of standardized beta-cell grafts in a liver segment of IDDM patients: graft and recipient characteristics in two cases of insulin-independence under maintenance immunosuppression for prior kidney graft.

Composition of human islet cell preparations for transplantation. Acta Diabetol. Anatomic and hormonal markers for the ventral primordium in the human pancreas.


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  • The pancreatic polypeptide-rich lobe of the human pancreas: definitive identification of its derivation from the ventral pancreatic primordium. The pancreatic polypeptide cells in the human pancreas: The effects of age and diabetes.

    Quantitative analysis of cell composition and purity of human pancreatic islet preparations

    Islet pathology and the pathogenesis of type 1 and type 2 diabetes mellitus revisited. Selective beta-cell loss and alpha-cell expansion in patients with type 2 diabetes mellitus in Korea. J Clin Endocrinol Metab. Beta-cell deficit and increased beta-cell apoptosis in humans with type 2 diabetes. Morphometry in endocrine pathology. Pathol Res Pract. The unique cytoarchitecture of human pancreatic islets has implications for islet cell function.

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    Assessment of human pancreatic islet architecture and composition by laser scanning confocal microscopy. J Histochem Cytochem. A novel method for the assessment of cellular composition and beta-cell viability in human islet preparations. Am J Transplant. A simple method of staining fresh and cultured islets. Use of oxygenated perfluorocarbon toward making every pancreas count. Automated method for isolation of human pancreatic islets. Islet transplantation in seven patients with type 1 diabetes mellitus using a glucocorticoid-free immunosuppressive regimen.

    Improved human islet isolation using a new enzyme blend, liberase. Weibel ER. Stereological Methods. London: Academic Press; Practical methods for biological morphometry. Islet isolation assessment in man and large animals. Acta Diabetol Lat. Endocytosis of low-density lipoprotein by human pancreatic beta cells and uptake in lipid-storing vesicles, which increase with age. Bonner-Weir S. Anatomy of the islets of Langerhans. In: Samols E, editor. The Endocrine Pancreas. New York: Raven Press; Cellular composition of the human diabetic pancreas. Quantitation of endocrine cell content in the pancreas of nondiabetic and diabetic humans.

    Dispersed adult rat pancreatic islet cells in culture: A, B, and D cell function. A method for the purification of single A, B, and D cells and for the isolation of coupled cells from isolated rat islets. Pipeleers DG. The biosociology of pancreatic B-cells.

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    Dean PM. Compensatory growth of pancreatic B-cells in adult rats after short-term glucose infusion. Changes in the size and number of secretion granules in the rat exocrine pancreas induced by feeding or stimulation in vitro. A morphometric study. Cell Tissue Res. Sato T, Herman L. Morphometry and elemental analysis of rat exocrine pancreas following administration of trypsin inhibitor. Acta Anat Basel ; 1 — Correlative morphometric and biochemical study on pancreatic amylase in normal and streptozotocin-diabetic rats.

    Metabolism of glucose in the islets of Langerhans. J Biol Chem. How does Europe PMC derive its citations network? Protein Interactions. Protein Families.

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    Nucleotide Sequences. Functional Genomics Experiments. Protein Structures. Gene Ontology GO Terms. Data Citations. Proteomics Data. Menu Formats. Full Text. Total tissue recovered in all fractions. Final islet pellet. Islet IC. Non-islet NIC. Total TC. Number of Total Cells n IE. Brissova et al. Keymeulen et al. Ichii et al. Street et al. Cabrera et al. Butler et al. Yoon et al. Sakuraba Mass culture of animal cell lines is fundamental to the manufacture of viral vaccines and other products of biotechnology.

    Culture of human stem cells is used to expand the number of cells and differentiate the cells into various somatic cell types for transplantation. Biological products produced by recombinant DNA rDNA technology in animal cell cultures include enzymes , synthetic hormones , immunobiologicals monoclonal antibodies , interleukins , lymphokines , and anticancer agents. Although many simpler proteins can be produced using rDNA in bacterial cultures, more complex proteins that are glycosylated carbohydrate-modified currently must be made in animal cells.

    An important example of such a complex protein is the hormone erythropoietin. The cost of growing mammalian cell cultures is high, so research is underway to produce such complex proteins in insect cells or in higher plants, use of single embryonic cell and somatic embryos as a source for direct gene transfer via particle bombardment, transit gene expression and confocal microscopy observation is one of its applications.

    It also offers to confirm single cell origin of somatic embryos and the asymmetry of the first cell division, which starts the process. Cell culture is also a key technique for cellular agriculture , which aims to provide both new products and new ways of producing existing agricultural products like milk, cultured meat , fragrances, and rhino horn from cells and microorganisms. It is therefore considered one means of achieving animal-free agriculture. It is also a central tool for teaching cell biology.

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    Research in tissue engineering , stem cells and molecular biology primarily involves cultures of cells on flat plastic dishes. This technique is known as two-dimensional 2D cell culture, and was first developed by Wilhelm Roux who, in , removed a portion of the medullary plate of an embryonic chicken and maintained it in warm saline for several days on a flat glass plate.

    From the advance of polymer technology arose today's standard plastic dish for 2D cell culture, commonly known as the Petri dish. Julius Richard Petri , a German bacteriologist , is generally credited with this invention while working as an assistant to Robert Koch.

    Various researchers today also utilize culturing laboratory flasks , conicals, and even disposable bags like those used in single-use bioreactors. Aside from Petri dishes, scientists have long been growing cells within biologically derived matrices such as collagen or fibrin, and more recently, on synthetic hydrogels such as polyacrylamide or PEG.

    They do this in order to elicit phenotypes that are not expressed on conventionally rigid substrates. There is growing interest in controlling matrix stiffness , [29] a concept that has led to discoveries in fields such as:. Cell culture in three dimensions has been touted as "Biology's New Dimension". Scaffold based cultures utilize an acellular 3D matrix or a liquid matrix.

    Scaffold-free methods are normally generated in suspensions. Eric Simon, in a NIH SBIR grant report, showed that electrospinning could be used to produced nano- and submicron-scale polystyrene and polycarbonate fibrous scaffolds specifically intended for use as in vitro cell substrates. It was noted that, as opposed to the flattened morphology typically seen in 2D culture, cells grown on the electrospun fibers exhibited a more histotypic rounded 3-dimensional morphology generally observed in vivo. As the natural extracellular matrix ECM is important in the survival, proliferation, differentiation and migration of cells, different hydrogel culture matrices mimicking natural ECM structure are seen as potential approaches to in vivo —like cell culturing.

    Several different types of hydrogels from natural and synthetic materials are available for 3D cell culture, including animal ECM extract hydrogels, protein hydrogels, peptide hydrogels, polymer hydrogels, and wood-based nanocellulose hydrogel. The magnetic nanoparticle assemblies consist of magnetic iron oxide nanoparticles, gold nanoparticles, and the polymer polylysine. Cell culture is a fundamental component of tissue culture and tissue engineering , as it establishes the basics of growing and maintaining cells in vitro.

    The major application of human cell culture is in stem cell industry, where mesenchymal stem cells can be cultured and cryopreserved for future use. Tissue engineering potentially offers dramatic improvements in low cost medical care for hundreds of thousands of patients annually. Vaccines for polio , measles , mumps , rubella , and chickenpox are currently made in cell cultures. Due to the H5N1 pandemic threat, research into using cell culture for influenza vaccines is being funded by the United States government.

    Novel ideas in the field include recombinant DNA -based vaccines, such as one made using human adenovirus a common cold virus as a vector, [53] [54] and novel adjuvants. Besides the culture of well-established immortalised cell lines, cells from primary explants of a plethora of organisms can be cultured for a limited period of time before sensecence occurs see Hayflick's limit.

    Cultured primary cells have been extensively used in research, as is the case of fish keratocytes in cell migration studies. Plant cell cultures are typically grown as cell suspension cultures in a liquid medium or as callus cultures on a solid medium. The culturing of undifferentiated plant cells and calli requires the proper balance of the plant growth hormones auxin and cytokinin. Cells derived from Drosophila melanogaster most prominently, Schneider 2 cells can be used for experiments which may be hard to do on live flies or larvae, such as biochemical studies or studies using siRNA.

    Cell lines derived from the army worm Spodoptera frugiperda , including Sf9 and Sf21 , and from the cabbage looper Trichoplusia ni , High Five cells , are commonly used for expression of recombinant proteins using baculovirus.

    For bacteria and yeasts, small quantities of cells are usually grown on a solid support that contains nutrients embedded in it, usually a gel such as agar, while large-scale cultures are grown with the cells suspended in a nutrient broth. The culture of viruses requires the culture of cells of mammalian, plant, fungal or bacterial origin as hosts for the growth and replication of the virus. Whole wild type viruses, recombinant viruses or viral products may be generated in cell types other than their natural hosts under the right conditions. Depending on the species of the virus, infection and viral replication may result in host cell lysis and formation of a viral plaque.

    From Wikipedia, the free encyclopedia. Main article: Cell isolation. Main article: List of contaminated cell lines. Main article: Passaging. Main articles: Transfection and Transformation genetics. Further information: Hybridoma. Main article: Plant tissue culture. See also: Tobacco BY-2 cells. Main article: Microbiological culture. Main article: Viral culture. This list is incomplete ; you can help by expanding it.

    Retrieved Archived from the original on Yale Alumni Magazine. Journal of Molecular and Cellular Cardiology. Selborne Biological Services. Fertility and Sterility. Cases of mistaken identity". Cancer Letters. International Journal of Cancer. Nature Reviews. Cell Notes. Archived from the original PDF on Human Reproduction Update. These systems are explored in the contexts of sociological importance, such as socialization, learning, gender roles, gender differences, sexuality, the family, deviance, and criminality. His research interests include any social-psychological topic that can be informed by biological concepts, particularly IQ and crime.

    Walsh is the author or coauthor of nine other books and over 60 journal articles or essays.