The NHLBI iPSC Core Facility started service in May 2011. The lab temporarily operated in 10/6N240 till relocation to 10/5N214 in May 2012. The major activities in the lab include lab set up, training, collaboration with outside groups, iPSC derivation for the Institute and standard iPSCs for other intramural institutes. (iPSC: induced Pluripotent Stem Cells) A. Lab Set up and Services 1. Equipment set up. The lab was opened for consultation in May 2011 and we started to purchase essential equipment from May till August 2011. 2. The current lab site 6N240 was made operational for research for research in May 2011, the initial renovation was finished by the end of July, and the lab was functional in mid August 2011. 3. The initial iPSC derivation was started in Dr. Boehms lab in July, and later moved to 6N240. We have generated more than 124 lines from 33 patient and control samples. 4. We set up the operational procedure to generate defined medium for iPSC maintenance and derivation. We have generated 200L medium. 5. We developed iPSC derivation procedures in defined medium with lentivirus, episomal and Sendai virus approaches. We also tested fibroblast, CD34+, peripheral blood cells, HUVEC and adipocyte. 6. In collaboration with NIH-CRM, we prepared three NIH control cell lines that will be distributed to researchers in the community as standard iPSC line. 7. We developed protocols to evaluate incubation conditions and quality control steps for new reagents. We also routinely screened our cell culture. 8. A standard protocol was set up to analyze stem cell nuclear and surface markers. We are now able to analyze multiple samples with multiple antibodies in a high throughput fashion. 9. In collaboration with the Transgenic Core, we currently have protocols to conduct teratoma formation assay on SCID mice;and teratoma is stained and analyzed by Pathology Core. B. Training activities We held 8 workshops on human ESC/iPSC culture techniques. We hosted 36 NHLBI researchers from 19 NHLBI labs and we also trained 13 researchers from other NIH institutes and outside groups. We also provided additional consultations to NIH research groups. C. Interaction with outside groups 1. Collaboration with NIH-CRM on control cell lines. 2. We are the founding member of the US Stem Cell Core Facility network. Invited Talks NIH-CRM, 01/17/2012 Defining Cell Culture Conditions for Translational Research: Starting from Human Pluripotent Stem Cells. NCI-Frederick, 01/26/2012, Optimizing Stem Cell Culture Conditions for Translational Research. Reviewer duty for peer-reviewed journals: PLOS One, Stem Cells   D. iPSC lines derived in the lab. We successfully derived >124 iPSCs from 33 patient or control samples using lentivirus, Sendai virus and episomal approach. These cells will be used for disease modeling and protocol development. E. Control cell lines for the NHLBI and NIH. We banked 9 cell lines as control lines for NIH researchers. These lines will be used to generate lineage tracing cells and can be used to standardize conditions and develop new protocols.

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Project End
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Budget End
Support Year
1
Fiscal Year
2012
Total Cost
$1,156,486
Indirect Cost
Name
National Heart, Lung, and Blood Institute
Department
Type
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Kaewkhaw, Rossukon; Swaroop, Manju; Homma, Kohei et al. (2016) Treatment Paradigms for Retinal and Macular Diseases Using 3-D Retina Cultures Derived From Human Reporter Pluripotent Stem Cell Lines. Invest Ophthalmol Vis Sci 57:ORSFl1-ORSFl11

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