Re: Farmas USA
OCAT Dicen por ahí que la emisión ya se ha completado con éxito. Luego acaban de encontrar una presentación de marzo de especulan que es la presentación a la FDA y con la que están exultantes. http://c.ymcdn.com/sites/casss.site-ym.com/resource/resmgr/Bioassays_Speaker_Slides/2015_BIOA_KlimanskayaIrina.pdf Copio uno de los mesajes de Ablidy, que sabe. Bioassays …..What were the data sets the could be use as in-process QC … gene expression ( PCR) and flow cytometry data cell surface markers…plus many picture from actual cells in vivo….. Why is this important when you are setting up a manufacturing process in biotech you better have purity , scalability, batch to batch reproducibility and in stem cells tolerance and potency….What have we learn most from Lanza ? Potency and how that is defined with tolerance for allogeneic cells that proliferate after differentiate into end results such as RPE to treat AMD/SMA /MMD for the masses b/c of manufacturability and cost ….less than $100 for under 300K dose product and you can sell it for $$$$$$$with very high profit margin’s Let’s look at Irina’s work of art in her presentation…..Remember she is the one to discover the RPE cell maturation process….. Slide # 2 You could find on your eye doctors wall in his waiting room as art autograph of RPE cells….. Ask the Dr. Ron types on the board….. Slide #3 Is plain and simple structure of the retina pay attention to the bottom of the slide Slides #4-5 Are visual physiology we have seen these slide before , but my guess is Irina was the original author Slide #5-6 Speaks for itself what happen to the dog for the patients ……what a beautiful marketing slide to get the non patient or investor to focus on the problem….. Slide # 9 Thesis of RPE Transplantation: Restoration of Anatomy & Function Slide #10 Generation of RPE from Human ES Cells with actual pictures of RPE cells that have Pigmented clusters are isolated Slide # 11-12 RPE Transdifferentiation in Culture with the RPE undergoes dedifferentiation during culture expansion and matures after reaching confluence in fresh cluster Slide #13 Post-Thaw Survival is Higher in Less Pigmented Cells what are you looking at on this slide ? Irina is showing batch to batch to the FDA …..as cell lines are pasted from one passage to the next Lanza has told us they can lose their potency b/ gene expression matures an culture conditions changes…..Irina shows the FDA they can start from scratch …..Ocata never need to get another RPE cell line ever !!!!!!!!!!!!!!!!!!! Slide # 14 Is Ocata ‘s in process early method strategy….. RPE Cells are Harvested Before They Fully Mature Slides #15-16 Is illustration the problems of 2D tissue cultures methods In Vitro vs. In Vivo Limitations Slide #16 This is an old slide form 2005, but is a great slide that gets your eye that is why I told Twenty2 to tweet this ….. RPE Multitasking Now we get to the meat and potatoes of her presentation to the FDA and their audience on bioassays Slides #17-20 Potency Assay for a Relevant Physiological Function of hESC-RPE….What is it and how does it work? Phagocytosis with the main function on a biologist could appreciate the elegant illustration and importance of engulfment, formation of phagosomes fusion of phagosomes with lysosomes…… Major point for the eye’s immune system antigen presentation to get rid of cell debris …..What is one of the functions of RPE CELLS? Slides # 21 Irina shows 1st Generation Potency Assay: Phagocytosis of Latex Beads by Electron Microscopynice but how could you use this in QC….Can not Slide # 22 the Home Run slide using Flow cytometry to show Current Phagocytosis Assay: Quantitative pHrodo-Bioparticle Uptake Let’s look at this slide more critical can you see the the resolution of the two histogram from the flow data ? Clears as day and night…Why the 37 vs. 4 degree from the control? You need heat tor energy to have high efficiency for phagocytosis…..Cells are slow at 4 degrees Slides #23-28 Irina explains why pHrodo Bioparticles Demonstrate the Completion of Several Phagocytosis Steps and is a bioassay … OS Phagocytis clearance: Recognition and Binding (OS is outer segments) She should has define this for the reader OS Phagocytosis clearance: Recognition and Binding what the PRE cells start the disease process/c incomplete OS digestion lipofuscin accumulation drusen formation retinal degeneration….Leading to Vision Diseases Related to Phagocytosis Defects Slide # 29 More flow data Vision Diseases Related to Phagocytosis Defects with the Phagocytosis Assay Using Bovine Photoreceptor OS – The Next Generation Irina needs to go to multi color analysis with flow …3-4 colors with different markers ….Why so the programmers can write software for hands off analysis for in process QC of Ocata manufacturing process Slide #30-36 Ocata trial results you all should know these slides Slides #37 1. RPE phagocytosis of photoreceptor outer segments (OS) is a physiologically relevant potency assay; QC friendly, relatively simple, and quantitative. 2. hESC-RPE are capable of OS phagocytosis and show normal progression through the stage of phagolysosome formation. 3. Mechanism of Action can be addressed by examining specific steps in the cascade of phagocytic events including: OS binding processing, and export. 1.How does the stage of maturity/pigmentation affect the phagocytosis rate? Multi color flow bioassays 2.Which rate of phagocytosis is physiologically appropriate? Determined by enzyme functions 3. Can we set quantitative parameters? Yes by using bead technology or qPCR 4. What is the OS phagocytosis capacity of hESC-RPE? This one only Lanza and Irina knows…. 5. is there OS clearance and lipofuscin accumulation in cultured RPE? Need a study …..if only the NIH had change the redefinition of pre embryonic stem cell to include MA-09 cell lines some hot shot PhD could be funded to do this study outside of the Ocata PRE project ….how FFB funding this basic research???? 6. How to correlate the in vitro data with anticipated in vivo performance of these cells? Proper clinical trials design……
«Después de nada, o después de todo/ supe que todo no era más que nada.»
