Pancreas organoids for type I diabetes mellitus - Is it feasible as a cell therapy?

Human pancreas organoids are a promising cell therapy candidate for Type I Diabetes Mellitus (T1DM). T1DM is a disabling chronic disease with a juvenile onset. Auto-immune destruction of pancreatic beta cells results in insufficient insulin production to manage blood glucose. This can lead, on the long-term, to serious complications, such as hypoglycemic episodes, neuropathy, retinopathy, and renal failure. The current standard of care is insulin therapy. However this cannot fully prevent long-term complications from arising, especially in patients suffering from uncontrolled T1DM with severe hypoglycemic episodes. Restoration of the natural blood glucose management can prevent complications, for instance by restoring the beta cell population with a cell therapy. Organoids, proclaimed by the journal Nature as “Method of the year 2017”, are 3D structures that can be generated from progenitor cells of many different organs, such as pancreas, liver, brain, lung, and heart. These miniature organs, when differentiated to produce insulin, can be a 3D cell therapy for T1DM. In case of the pancreas, these can be generated from adult-derived progenitor cells, which have a safer profile than ESC or iPSC based cell therapies for T1DM. Lonza Netherlands is a partner within the LSFM4LIFE project: a European consortium of universities and industrial partners that aims to produce a GMP batch of human pancreas organoids. To reach this goal, two milestones need to be achieved. First, to transfer the research process of organoid production into GMP. Specific challenges are a scalable GMP compliant platform for 3D culture and replacing research materials (e.g. complex media formulations and culture substrates) with GMP compliant substitutes. Additionally in process controls and QC assays are in its infancy for organoids and need to be developed. The second milestone is to develop a strategy for commercialization of this therapy, for which a number of analyses have been performed. A market and SWOT analysis have been performed (Figure 1). A clinical strategy is proposed to first access the market with an introduction into a small patient cohort and then to stay on the market by reaching all T1DM patients. An assessment of the manufacturing cost of goods is made for the current process, as well as how it could be envisioned in a commercial setting. From these costs, the cost-effectiveness compared to insulin therapy and islet of Langerhans transplantation is evaluated [1]. In conclusion, while the potential for organoids as a cellular therapy is considerable, this paper addresses the progress so far and the major challenges ahead. Please click Additional Files below to see the full abstract

Design and functional testing of a multichamber perfusion platform for three-dimensional scaffolds

Perfusion culture systems are widely used in tissue engineering applications for enhancing cell culture viability in the core of three-dimensional scaffolds. In this work, we present a multichamber confined-flow perfusion system, designed to provide a straightforward platform for three-dimensional dynamic cell cultures. The device comprises 6 culture chambers allowing independent and simultaneous experiments in controlled conditions. Each chamber consists of three parts: a housing, a deformable scaffold-holder cartridge, and a 7 mL reservoir, which couples water-tightly with the housing compressing the cartridge. Short-term dynamic cell seeding experiments were carried out with MC3T3-E1 cells seeded into polycaprolactone porous scaffolds. Preliminary results revealed that the application of flow perfusion through the scaffold favored the penetration of the cells to its interior, producing a more homogeneous distribution of cells with respect to dropwise or injection seeding methods. The culture chamber layout was conceived with the aim of simplifying the user operations under laminar flow hood and minimizing the risks for contamination during handling and operation. Furthermore, a compact size, a small number of components, and the use of bayonet couplings ensured a simple, fast, and sterility-promoting assembling. Finally, preliminary in vitro tests proved the efficacy of the system in enhancing cell seeding efficiency, opening the way for further studies addressing long-term scaffold colonization

Fabrication of 3D cell-laden hydrogel microstructures through photo-mold patterning

Native tissues are characterized by spatially organized three-dimensional (3D) microscaled units which functionally define cells–cells and cells–extracellular matrix interactions. The ability to engineer biomimetic constructs mimicking these 3D microarchitectures is subject to the control over cell distribution and organization. In the present study we introduce a novel protocol to generate 3D cell laden hydrogel micropatterns with defined size and shape. The method, named photo-mold patterning (PMP), combines hydrogel micromolding within polydimethylsiloxane (PDMS) stamps and photopolymerization through a recently introduced biocompatible ultraviolet (UVA) activated photoinitiator (VA-086). Exploiting PDMS micromolds as geometrical constraints for two methacrylated prepolymers (polyethylene glycol diacrylate and gelatin methacrylate), micrometrically resolved structures were obtained within a 3 min exposure to a low cost and commercially available UVA LED. The PMP was validated both on a continuous cell line (human umbilical vein endothelial cells expressing green fluorescent protein, HUVEC GFP) and on primary human bone marrow stromal cells (BMSCs). HUVEC GFP and BMSCs were exposed to 1.5% w/v VA-086 and UVA light (1 W, 385 nm, distance from sample = 5 cm). Photocrosslinking conditions applied during the PMP did not negatively affect cells viability or specific metabolic activity. Quantitative analyses demonstrated the potentiality of PMP to uniformly embed viable cells within 3D microgels, creating biocompatible and favorable environments for cell proliferation and spreading during a seven days' culture. PMP can thus be considered as a promising and cost effective tool for designing spatially accurate in vitro models and, in perspective, functional constructs

Embryoid body size-mediated differential endodermal and mesodermal differentiation using polyethylene glycol (PEG) microwell array

Embryoid bodies have a number of similarities with cells in gastrulation, which provides useful biological information about embryonic stem cell differentiation. Extensive research has been done to study the control of embryoid body-mediated embryonic stem cell differentiation in various research fields. Recently, microengineering technology has been used to control the size of embryoid bodies and to direct lineage specific differentiation of embryonic stem cells. However, the underlying biology of developmental events in the embryoid bodies of different sizes has not been well elucidated. In this study, embryoid bodies with different sizes were generated within microfabricated PEG microwell arrays, and a series of gene and molecular expressions related to early developmental events was investigated to further elucidate the size-mediated differentiation. The gene and molecular expression profile suggested preferential visceral endoderm formation in 450 μm embryoid bodies and preferential lateral plate mesoderm formation in 150 μm embryoid bodies. These aggregates resulted in higher cardiac differentiation in 450 μm embryoid bodies and higher endothelial differentiation in 150 μm embryoid bodies, respectively. Our findings may provide further insight for understanding embryoid body size-mediated developmental progress.National Science Foundation (U.S.) (CAREER Award DMR0847287)United States. Office of Naval Research (Naval Research Young National Investigator Award)National Institutes of Health (U.S.) (HL092836, EB02597, AR057837

Strategic Structure for Organizational Performance (A Case Study: Leather Industry)

ABSTRACT Structure management is defined as the set of decisions and actions resulting in formulation and implementation of strategies designed to achieve the objectives of an organization that it involves after the super ordinate goal and strategy. In order to performance increase of organizations because of competition conditions in nowadays world with more various threats, perform of necessary actions is required. Therefore, the proposed model including creates structure, balancer, execution and supporter with the name of organizational strategic structure examined and tested in six industrial organizations such as samples. For this reason, this article is used data collection from studies in six industrial organizations in North West of Iran that the main purpose of it was to examine the organizational performance and strategic structure. The examples have clear implications for practitioners for improve organizational performance as far as possible via strategic structure management. The final results showed that organizational performance quantity in compliance with determined criteria's of evaluation at the time of before and after perform of suggested model for each organization are deferent. The other implication of the research is that strategic structure impacts on organizational performance. Further, results showed that if the total average values of each worker be very high, the create structure will be suitable for him, but if such values be very low, the execution place in Strategic structure will be proposed. In other wise, if the total average values of person be medium, he or she will put in balancer or supporting place in strategic structure. Any how, in accordance with organizational performance value increase at other organizations that strategic structure model performed more than about 70 percent, results showed that upper than this range, the positive change of performance value will be possible in organization

Design and Functional Testing of a Multichamber Perfusion Platform for Three-Dimensional Scaffolds

Perfusion culture systems are widely used in tissue engineering applications for enhancing cell culture viability in the core of three-dimensional scaffolds. In this work, we present a multichamber confined-flow perfusion system, designed to provide a straightforward platform for three-dimensional dynamic cell cultures. The device comprises 6 culture chambers allowing independent and simultaneous experiments in controlled conditions. Each chamber consists of three parts: a housing, a deformable scaffold-holder cartridge, and a 7 mL reservoir, which couples water-tightly with the housing compressing the cartridge. Short-term dynamic cell seeding experiments were carried out with MC3T3-E1 cells seeded into polycaprolactone porous scaffolds. Preliminary results revealed that the application of flow perfusion through the scaffold favored the penetration of the cells to its interior, producing a more homogeneous distribution of cells with respect to dropwise or injection seeding methods. The culture chamber layout was conceived with the aim of simplifying the user operations under laminar flow hood and minimizing the risks for contamination during handling and operation. Furthermore, a compact size, a small number of components, and the use of bayonet couplings ensured a simple, fast, and sterility-promoting assembling. Finally, preliminary in vitro tests proved the efficacy of the system in enhancing cell seeding efficiency, opening the way for further studies addressing long-term scaffold colonization