SBIR-STTR Award

Conventional culture vessels are not designed for physiological oxygen delivery. Both hyperoxia and hypoxia -commonly observed when culturing cells and tissues in regular plastic ware- have been linked to reduced cellular function and death. This is particularly true of pancreatic beta cells, which are highly sensitive to sub- and super-physiological oxygen concentrations. We have addressed this problem by devising a novel culture device, the "oxygen sandwich". This simple system is designed to deliver oxygen in a physiological-like fashion by means of a basal air-permeable perfluorocarbon-silicone (PFC/Si) membrane. Our long-term goal is to establish this system as the new culture standard for beta cell biology applications, a rapidly expanding market that includes in vitro research on adult/fetal islets of any species, as well as beta cell differentiation studies. In this context, the specific aim of this Phase I proposal is to establish proof of concept that the enhanced in vitro survival and function observed in PFC/Si-cultured islets results in better pre-clinical transplantation outcomes. Our research design is based on a marginal mass xenotransplantation model (human islets into diabetic nu/nu immunodeficient mice), where the potential benefits of PFC/Si culture prior to transplantation should translate into a shorter time to diabetes reversal compared to animals transplanted with control (regular culture) islets. Our initial results are strongly supportive of the feasibility of this proposal, and we have partnered with the Diabetes Research Institute at the University of Miami to accomplish our goals in a timely manner. Success in our research would fill a widely acknowledged gap in our ability to preserve islet cell function and survival in vitro, confirming this system as a potential new standard for beta cell biology and differentiation studies. Such positive outcome would be highly relevant to the mission of the National Institutes of Diabetes and Digestive and Kidney Diseases (NIDDK), and might ultimately be of clinical significance for human islet/beta cell transplantation.

Public Health Relevance:
Beta cell research is a large field where basic science interest is further fueled by current (islet transplantation) and prospective (stem cells and xenotransplantation) clinical therapies for diabetes. However, beta cell studies are compromised by the inability of conventional culture systems to provide physiological oxygenation. Since our proposal aims at testing a novel culture device specifically designed to overcome this problem, these studies are highly relevant to the mission of the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) and may ultimately have a significant impact on public health.

Public Health Relevance Statement:
NARRATIVE Beta cell research is a large field where basic science interest is further fueled by current (islet transplantation) and prospective (stem cells and xenotransplantation) clinical therapies for diabetes. However, beta cell studies are compromised by the inability of conventional culture systems to provide physiological oxygenation. Since our proposal aims at testing a novel culture device specifically designed to overcome this problem, these studies are highly relevant to the mission of the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) and may ultimately have a significant impact on public health.

NIH Spending Category:
Autoimmune Disease; Bioengineering; Biotechnology; Diabetes; Regenerative Medicine; Transplantation

Project Terms:
2-Deoxy-2-((methylnitrosoamino)carbonyl)amino-D-glucose; 2-deoxy-2-(3-methyl-3-nitrosoureido)-D-glucopyranose; 2-deoxy-2-[[(methylnitrosamino)-carbonyl]amino]-D-glucopyranose; 21+ years old; Academia; Address; Adult; Air; Animals; Anoxia; Articulation; Arts; Autoregulation; B9 endocrine pancreas; Basic Research; Basic Science; Beta Cell; Biological Models; Biology; Blood flow; Bone Marrow; Cardiac Myocytes; Cardiocyte; Cell Count; Cell Culture Techniques; Cell Differentiation; Cell Differentiation process; Cell Function; Cell Line; Cell Lines, Strains; Cell Number; Cell Process; Cell Survival; Cell Therapy; Cell Viability; Cell physiology; CellLine; Cells; Cellular Function; Cellular Physiology; Cellular Process; Cellular biology; Cessation of life; Clinical; Clinical Trials, Phase II; Culture Media; Custom; D-Glucose; Data; Death; Development; Devices; Dextrose; Diabetes Mellitus; Diabetic mouse; Diffusion; Discipline; Ensure; Equilibrium; Equipment; Explosion; Fluorocarbons; Future; Gastrointestinal Tract, Pancreas; Generalized Growth; Glucose; Goals; Grafting, Islets of Langerhans; Growth; Heart myocyte; Hepatic Cells; Hepatic Parenchymal Cell; Hepatocyte; Heterograft; Homeostasis; Human; Human, Adult; Human, General; Hyperoxia; Hypoxia; Hypoxic; Immunocompromised; Immunocompromised Host; Immunocompromised Patient; Immunodeficient Mouse; Immunosuppressed Host; In Vitro; Individual; Industry; Insulin Cell; Insulin Secreting Cell; Intervention; Intervention Strategies; Investigators; Islands of Langerhans; Islet Cell; Islet Cells; Islets of Langerhans; Islets of Langerhans Transplantation; Joints; Knowledge; Laboratories; Legal patent; Length; Link; Liver Cells; Man (Taxonomy); Man, Modern; Marketing; Measures; Medical; Membrane; Methods; Mission; Model System; Modeling; Models, Biologic; Molecular Biology Techniques; Mother Cells; Muscle Cells, Cardiac; Muscle Cells, Heart; Myocytes, Cardiac; NIDDK; National Institute of Diabetes and Digestive and Kidney Diseases; National Institute of Digestive Diseases and Kidney Disorders; Nesidioblasts; O element; O2 element; Organ; Outcome; Oxygen; Oxygen Deficiency; Pancreas; Pancreas, Endocrine; Pancreatic; Pancreatic Islets; Pancreatic beta Cell; Pars endocrina pancreatis; Patents; Perfluorocarbons; Phase; Phase 2 Clinical Trials; Phase II Clinical Trials; Physicians; Physiologic; Physiological; Physiological Homeostasis; Plastics; Preparation; Process; Production; Progenitor Cells; Public Health; Qualifying; Research; Research Design; Research Institute; Research Personnel; Researchers; Reticuloendothelial System, Bone Marrow; Role; STZ; Silicones; Stem Cell Research; Stem cells; Streptozocin; Streptozotocin; Structure of beta Cell of islet; Study Type; Subcellular Process; System; System, LOINC Axis 4; Technology; Temperature; Testing; Therapy, Cell; Time; Tissue Growth; Training; Translating; Translatings; Translational Research; Translational Research Enterprise; Translational Science; Transplantation; Transplantation, Heterologous; Transplantation, Islands of Langerhans; Transplantation, Islands of Pancreas; Transplantation, Islet; Transplantation, Pancreatic Islets; Universities; Variant; Variation; Xenograft; Xenograft procedure; Xenotransplantation; Zanosar; adult human (21+); balance; balance function; base; beta cell development; cardiomyocyte; cell biology; cell-based therapy; clinical significance; clinically significant; commercialization; cultured cell line; design; designing; diabetes; diabetes mellitus therapy; diabetes therapy; diabetic; endocrine pancreas; endocrine pancreas development; experience; feeding; fetal; growth media; immortalized cell; immunosuppressed patient; improved; interest; interventional strategy; islet; islet beta cell transplantation; islet cell transplant; islet cell transplantation; islet development; islet progenitor; islet stem cells; islet transplantation; language translation; membrane structure; mouse model of diabetes; multidisciplinary; novel; ontogeny; oxygen stress (breathing); pancreas beta cell; pancreas stem cells; phase 2 study; phase 2 trial; phase II trial; pre-clinical; preclinical; prevent; preventing; prospective; protocol, phase II; prototype; public health medicine (field); public health relevance; social role; stem cell differentiation; study design; study, phase II; success; tissue culture; tissue/cell culture; tool; translation research enterprise; transplant

Conventional culture vessels are not designed for physiological oxygen delivery. Both hyperoxia and hypoxia - commonly observed when culturing cells and tissues in regular plasticware- have been linked to reduced cellular function and death. An adequate means to provide oxygenation is also critical for stem cell applications in which the differentiation outcome is dependent on oxygen tension levels. We have addressed this problem by devising a novel culture device, the "oxygen sandwich". This simple system is designed to deliver oxygen in a quasi-physiological fashion by means of a basal air-permeable perfluorocarbon-silicone (PFC/Si) membrane. Our long-term goal is to establish this system as a new standard for tissue culture, both for applications in which oxygen diffusion rates become limiting (such as 3D culture) and for those that require precise adjustments of oxygenation to steer stem cell differentiation in the desired direction. We will focus our proof-of- concept work on islet/beta cell biology. This is a rapidly expanding market that includes clinical uses (islet transplantation), active in vitro research on adult/fetal islets of several species, and pre-clinicl stem cell research with the potential to revolutionize the treatment of diabetes within the next 5-10 years. The pertinence of our model choice is highlighted by two well-documented observations: [1] Pancreatic beta cells (the current end product used in clinical therapies for diabetes) are highly sensitive to sub- and super-physiological oxygen concentrations; and [2] Stem cell differentiation into beta cells (the subject of worldwide research to replace cadaveric islets for future clinical uses) is exquisitely dependent on evolving oxygen tensions. Our Phase I studies aimed at demonstrating that the enhanced in vitro survival and function observed in PFC/Si-cultured islets also resulted in better pre-clinical transplantation outcomes using a marginal mass xenotransplantation model (human islets into diabetic nu/nu immunodeficient mice). After the successful completion of these studies, our Phase II proposal is based on the following specific aims: (1) Scaling-up and definition of manufacturing process (QA & QC, sterilization) for mass production of PFC/Si culture devices; and (2) biological testing and reproducibility studies in human islets and embryonic stem cells (hESc). This application benefits from the assembly of first-rate teams with highly complementary expertise. Our Phase I results are strongly supportive of the feasibility of this proposal. Success in our research would fill a widely acknowledged gap in our ability to preserve islet cell function and survival in vitro confirming this system as a potential new standard for beta cell biology and differentiation studies. As such positive outcome might ultimately speed up the applicability of new-generation beta cell replacement therapies, this project is greatly relevant to the mission of the National Institutes of Diabetes and Digestive and Kidney Diseases (NIDDK). Success in these studies will also provide proof of principle of the superiority of our oxygen enhancing technology for many other cell culture applications.

Public Health Relevance Statement:
: NARRATIVE Beta cell research is a broad field where basic science interest is further fueled by current (islet transplantation) and prospective (stem cells and xenotransplantation) clinical therapies for diabetes. However, beta cell studies are compromised by the inability of conventional culture systems to provide physiological oxygenation. Since our proposal aims at testing a novel culture device specifically designed to overcome this problem, these studies are highly relevant to the mission of the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK). Many other cell culture applications may also directly benefit from the development of this technology. The successful completion of our studies, therefore, may ultimately have a significant impact on public health.

Project Terms:
Academia; Address; Administrator; Adult; Air; Award; base; Basic Science; Beta Cell; beta cell replacement; Biological Testing; Biology; Blood flow; California; Cell Culture Techniques; Cell Death; Cell Differentiation process; Cell physiology; Cell Survival; Cell Therapy; cell type; Cells; Cellular biology; Cessation of life; Clinical; commercialization; Custom; design; Development; Devices; Diabetes Mellitus; diabetes mellitus therapy; diabetic; Diffusion; Digestive System Disorders; Discipline; embryonic stem cell; Employee Strikes; Ensure; Equipment; Evolution; experience; fetal; flasks; Fluorocarbons; Funding; Future; Generations; Goals; Growth; Human; Hyperoxia; Hypoxia; Immunodeficient Mouse; improved; In Vitro; Industry; Institutes; Insulin-Dependent Diabetes Mellitus; interest; islet; Islet Cell; islet stem cells; Islets of Langerhans; Islets of Langerhans Transplantation; Kidney Diseases; Knowledge; Laboratories; Legal patent; Link; manufacturing process; Marketing; Medical; Membrane; Methods; Mission; Modeling; novel; Organ; Outcome; Oxygen; Oxygen measurement, partial pressure, arterial; Pancreas; Pattern; Phase; phase 1 study; Physicians; Physiological; pre-clinical; prevent; Production; prospective; public health medicine (field); Regenerative Medicine; Replacement Therapy; Reproducibility; Research; Research Institute; scale up; Silicones; Speed (motion); stem cell differentiation; Stem Cell Research; stem cell therapy; Stem cells; Sterilization; Structure of beta Cell of islet; success; System; Technology; technology development; Temperature; Testing; tissue culture; tissue/cell culture; Tissues; tool; Translational Research; Translations; Transplantation; trend; Universities; Washington; Work; Xenograft procedure