A Scalable System for Production of Functional Pancreatic Progenitors from Human Embryonic Stem Cells

Development of a human embryonic stem cell (hESC)-based therapy for type 1 diabetes will require the translation of proof-of-principle concepts into a scalable, controlled, and regulated cell manufacturing process. We have previously demonstrated that hESC can be directed to differentiate into pancreatic progenitors that mature into functional glucose-responsive, insulin-secreting cells in vivo. In this study we describe hESC expansion and banking methods and a suspension-based differentiation system, which together underpin an integrated scalable manufacturing process for producing pancreatic progenitors. This system has been optimized for the CyT49 cell line. Accordingly, qualified large-scale single-cell master and working cGMP cell banks of CyT49 have been generated to provide a virtually unlimited starting resource for manufacturing. Upon thaw from these banks, we expanded CyT49 for two weeks in an adherent culture format that achieves 50–100 fold expansion per week. Undifferentiated CyT49 were then aggregated into clusters in dynamic rotational suspension culture, followed by differentiation en masse for two weeks with a four-stage protocol. Numerous scaled differentiation runs generated reproducible and defined population compositions highly enriched for pancreatic cell lineages, as shown by examining mRNA expression at each stage of differentiation and flow cytometry of the final population. Islet-like tissue containing glucose-responsive, insulin-secreting cells was generated upon implantation into mice. By four- to five-months post-engraftment, mature neo-pancreatic tissue was sufficient to protect against streptozotocin (STZ)-induced hyperglycemia. In summary, we have developed a tractable manufacturing process for the generation of functional pancreatic progenitors from hESC on a scale amenable to clinical entry

Measurement of direct photon production in p + p collisions at sqrt(s) = 200 GeV

Cross sections for mid-rapidity production of direct photons in p+p collisions at the Relativistic Heavy Ion Collider (RHIC) are reported for 3 < p_T < 16 GeV/c. Next-to-leading order (NLO) perturbative QCD (pQCD) describes the data well for p_T > 5 GeV/c, where the uncertainties of the measurement and theory are comparable. We also report on the effect of requiring the photons to be isolated from parton jet energy. The observed fraction of isolated photons is well described by pQCD for p_T > 7 GeV/c.Comment: 330 authors, 6 pages text, 3 figures, one table. Submitted to Phys. Rev. Lett. Plain text data tables for the points plotted in figures for this and previous PHENIX publications are (or will be) publicly available at http://www.phenix.bnl.gov/papers.htm

Elliptic flow for phi mesons and (anti)deuterons in Au+Au collisions at sqrt(s_NN) = 200 GeV

Differential elliptic flow (v_2) for phi mesons and (anti)deuterons (d^bar)d is measured for Au+Au collisions at sqrt(s_NN) = 200 GeV. The v_2 for phi mesons follows the trend of lighter pi^+/- and K^+/- mesons, suggesting that ordinary hadrons interacting with standard hadronic cross sections are not the primary driver for elliptic flow development. The v_2 values for (d^bar)d suggest that elliptic flow is additive for composite particles. This further validation of the universal scaling of v_2 per constituent quark for baryons and mesons suggests that partonic collectivity dominates the transverse expansion dynamics.Comment: 343 authors, 6 pages text, 3 figures, REVTeX4. To be submitted to Physical Review Letters. Plain text data tables for the points plotted in figures for this and previous PHENIX publications are (or will be) publicly available at http://www.phenix.bnl.gov/papers.htm

J/psi production from proton-proton collisions at sqrt(s) = 200 GeV

J/psi production has been measured in proton-proton collisions at sqrt(s)= 200 GeV over a wide rapidity and transverse momentum range by the PHENIX experiment at RHIC. Distributions of the rapidity and transverse momentum, along with measurements of the mean transverse momentum and total production cross section are presented and compared to available theoretical calculations. The total J/psi cross section is 3.99 +/- 0.61(stat) +/- 0.58(sys) +/- 0.40(abs) micro barns. The mean transverse momentum is 1.80 +/- 0.23(stat) +/- 0.16(sys) GeV/c.Comment: 326 authors, 6 pages text, 4 figures, 1 table, RevTeX 4. To be submitted to PRL. Plain text data tables for the points plotted in figures for this and previous PHENIX publications are (or will be) publicly available at http://www.phenix.bnl.gov/papers.htm

Centrality Dependence of Charm Production from Single Electrons in Au+Au Collisions at sqrt(s_NN) = 200 GeV

The PHENIX experiment has measured mid-rapidity transverse momentum spectra (0.4 < p_T < 4.0 GeV/c) of single electrons as a function of centrality in Au+Au collisions at sqrt(s_NN) = 200 GeV. Contributions to the raw spectra from photon conversions and Dalitz decays of light neutral mesons are measured by introducing a thin (1.7% X_0) converter into the PHENIX acceptance and are statistically removed. The subtracted ``non-photonic'' electron spectra are primarily due to the semi-leptonic decays of hadrons containing heavy quarks (charm and bottom). For all centralities, charm production is found to scale with the nuclear overlap function, T_AA. For minimum-bias collisions the charm cross section per binary collision is N_cc^bar/T_AA = 622 +/- 57 (stat.) +/- 160 (sys.) microbarns.Comment: 326 authors, 4 pages text, 3 figures, 1 table, RevTeX 4. To be submitted to Physical Review Letters. Plain text data tables for the points plotted in figures for this and previous PHENIX publications are (or will be) publicly available at http://www.phenix.bnl.gov/papers.htm

Functional Beta Cell Mass from Device-Encapsulated hESC-Derived Pancreatic Endoderm Achieving Metabolic Control

Summary: Human stem cells represent a potential source for implants that replace the depleted functional beta cell mass (FBM) in diabetes patients. Human embryonic stem cell-derived pancreatic endoderm (hES-PE) can generate implants with glucose-responsive beta cells capable of reducing hyperglycemia in mice. This study with device-encapsulated hES-PE (4 × 106 cells/mouse) determines the biologic characteristics at which implants establish metabolic control during a 50-week follow-up. A metabolically adequate FBM was achieved by (1) formation of a sufficient beta cell number (>0.3 × 106/mouse) at >50% endocrine purity and (2) their maturation to a functional state comparable with human pancreatic beta cells, as judged by their secretory responses during perifusion, their content in typical secretory vesicles, and their nuclear NKX6.1-PDX1-MAFA co-expression. Assessment of FBM in implants and its correlation with in vivo metabolic markers will guide clinical translation of stem cell-derived grafts in diabetes. : In this article, Pipeleers and colleagues demonstrate that subcutaneous implants of device-encapsulated human stem cell-derived pancreatic endoderm can generate a functional beta cell mass that establishes sustained glucose control in mice. They identified their biologic characteristics and correlation with in vivo outcome. Data and methods are expected to guide clinical translation to beta cell replacement therapy in diabetes. Keywords: stem cell-derived pancreatic endoderm, stem cell therapy, diabetes, encapsulation, differentiation, functional maturation, functional beta cell mass, metabolic contro

Hugo

Our earlier robots have not solved the Field Robot Event’s row-following problem with a sufficient degree of robustness. The objective of the work presented here was to build a robot that can detect rows consisting of small or large maize plants by using a camera system; and to provide this robot with robust localization and navigation by using probabil-istic methods to process the data from the vision system in conjunction with data from other sensors. We employed a particle filter approach where information from the robot’s wheel encoders and a gyroscope is used in the control step and where the filter is updated using information from a downward-looking camera and a laser scanner. For the weed detection and control tasks, the robot is equipped with a self-contained spray unit consist-ing of two CMUCAM3 camera’s and four narrow-cone nozzles (two on each side of the robot) which allow for precision-treatment of small areas. At the Field Robot Event, the robot was able to follow rows and turn into the correct new row in all tasks. No manual intervention was necessary; the first objective was met. In the days and weeks leading up to the event, it was demonstrated that the robot can navigate even when the maize plants are very small. Thus, the second objective was also met. However, weed detection was less than perfect. It turned out to be more sensitive to the light conditions than we had realized. Also, the turf patches were placed almost between the maize plants instead of well inside the row, and were out of the camera’s view. In conclusion, the robot is capable of a high degree of autonomy in the tasks of the Field Robot Event: it didn’t once get lost and it damaged few plant

Immunogenicity of human embryonic stem cell-derived beta cells

Therapeutic cell differentiatio