Lab Relocation and Services (May 2012 April 2013) 1. The lab was moved from 10/6N240 to 10/5N214 in May 2012, and the services resume in June 2012. 2. The lab encountered multiple incidences related to the malfunction of Nitrogen and CO2 supply, and a few rounds of experiments were severely afffected. New set of liquid Nitrogen and CO2 tank system was installed for cell culture room in September 2012, and significant improvement has been observed. 3. The lab personnel raised health concerns related to uncomfortable work environment due to equipment noises in the lab, and a report was sent to the biosafety committee. As a result, extra space was assigned as equipment room, which will be available in 6 months. 4. In collaboration with NIH-CRM and Manfred Boehm lab, we expanded and distributed additional NIH control iPSC lines. 5. We generated more than 200 iPSC lines from 40 patient samples from NHLBI and multiple other NIH institutes. 6. We developed a semi-high throughput platform to derivate iPSCs from 24 patient samples simultaneously, and we were able to obtain iPSCs from 23 out of 24 patient samples. Further improvement is ongoing. 7. We developed new protocols to differentiate iPSCs as embryoid bodies. Paul Hwang, Cindy Dunbar and Haiming Caos labs are using it successfully. 8. We are developing new protocols to differentiate iPSCs to cardiomyocytes and hepatocytes in defined monolayer culture. Further improvement is underway. When the procedure is finished, we hope to have the capacity to supply cardiomyocyte, hepatocytes and progenitor sources for the whole Institute. 9. We set up in-house TALEN assembly procedure for genomic engineering. In initial test, we generated TALEN pairs for Toren Finkel and Manfred Boehm labs. 10. We started to use iLab system to document and manage the Core services in February 2013, and will fully implement it the end of 2013. Training activities In the last year, we held one group and 8 individual workshops on human ESC/iPSC culture methods. We also provided consultations to various NIH research groups. Six presentations were made to promote iPSC related research: Center for Molecular Medicine, Hematology Branch, Adelstein Lab, Zhao Lab, Leonard Lab and NCATS. Participation of grant applications and collaborations 1. Tobacco Regulatory Science Award: Boehm/Finkel (NHLBI);Gerhold/McKew (NCATS);Rao (NIH-CRM). 2. Bench to Bedside Cardiomyocyte based disease modeling: Finkel (NHLBI);McAreavey (NIH CC), Barac (Georgetown). Submitted. 3. NIH-CRM iPS derived vascular progenitors for the treatment of heart failure: Boehm/Horvath (NHLBI);Robey(NIDCR). No fund this year. 4. NIH-CRM iPSC Core Grant. No fund this year. 5. LINCS L1000 project with Broad Institute will move forward. 6. Cell therapy trial to test autologous iPSC-derived cardiomyocyte progenitor cells for the treatment of ischemic heart disease Boehm (NHBLI), Horvath (NHLBI), Murry (U. Washington) LOI submitted. 7. iPSC-derived Human Hepatocytes: A Treatment Model for End-Stage and Genetic Liver Diseases - Liang (NIDDK), Shamburek (NHLBI), Dunbar (NHLBI), Stronceck (CC), Liu (NHGRI) LOI submitted. Interaction with outside groups 8. Collaboration with NIH-CRM on control cell lines. 9. A founding member for the National Stem Cell Core Facility Group. 10. Collaboration with NIST to standardize stem cell imaging techniques. Invited Talks NIH-CRM Stem Cell Research Symposium, 05/10/2012 Control iPSC lines and Method Standardization. NIST, 06/15/2012, Defining Cell Culture Conditions for Translational Research: Starting from Human Pluripotent Stem Cells. Translational Science Mini-Symposium, 05/02/2013, Human Pluripotent Stem Cell Derivation and Differentiation for Translational Research Poster Presentations Maryland Stem Cell Foundation 2012 Keystone Symposium, February 2013 NIH Out Research Activities Mountain Vista Governors School in Warrenton, VA: Lab tour and PowerPoint presentation. October 26, 2012 Candidate site for Congressional and Departmental site visits. Reviewer duty for peer-reviewed journals: PLoS One, Stem Cells, Cancer, International Journal of Biological Sciences Publications citing the services: 1. Beers J, Gulbranson DR, George N, Siniscalchi LI, Jones J, Thomson JA and Chen G. Passaging and colony expansion of human pluripotent stem cells by enzyme-free dissociation in chemically defined culture conditions. Nature Protocols, 2012 2. Chen KG, Mallon BS, Hamilton RS, Kozhich OA, Park K, Hoeppner DJ, Robey PG, McKay RD. Non-colony type monolayer culture of human embryonic stem cells.
Stem Cell Res. 2012. 3. Chen G. Cryopreservation of human pluripotent stem cells in defined conditions. StemBook, 2012. 4. Chen G. iPSC Derivation from fibroblast in chemically defined medium. StemBook, 2012. 5. Chen G. Splitting hESC/hiPSC lines with EDTA in feeder free conditions. StemBook, 2012.

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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
Lin, Yongshun; Linask, Kaari L; Mallon, Barbara et al. (2016) Heparin Promotes Cardiac Differentiation of Human Pluripotent Stem Cells in Chemically Defined Albumin-Free Medium, Enabling Consistent Manufacture of Cardiomyocytes. Stem Cells Transl Med :
Hong, So Gun; Lin, Yongshun; Dunbar, Cynthia E et al. (2016) The Role of Nonhuman Primate Animal Models in the Clinical Development of Pluripotent Stem Cell Therapies. Mol Ther 24:1165-9
Qin, Haiying; Cho, Monica; Haso, Waleed et al. (2015) Eradication of B-ALL using chimeric antigen receptor-expressing T cells targeting the TSLPR oncoprotein. Blood 126:629-39
Cerbini, Trevor; Luo, Yongquan; Rao, Mahendra S et al. (2015) Transfection, selection, and colony-picking of human induced pluripotent stem cells TALEN-targeted with a GFP gene into the AAVS1 safe harbor. J Vis Exp :
Kaewkhaw, Rossukon; Kaya, Koray Dogan; Brooks, Matthew et al. (2015) Transcriptome Dynamics of Developing Photoreceptors in Three-Dimensional Retina Cultures Recapitulates Temporal Sequence of Human Cone and Rod Differentiation Revealing Cell Surface Markers and Gene Networks. Stem Cells 33:3504-18
Zou, Donghua; McSweeney, Colleen; Sebastian, Aswathy et al. (2015) A critical role of RBM8a in proliferation and differentiation of embryonic neural progenitors. Neural Dev 10:18
Cerbini, Trevor; Funahashi, Ray; Luo, Yongquan et al. (2015) Transcription activator-like effector nuclease (TALEN)-mediated CLYBL targeting enables enhanced transgene expression and one-step generation of dual reporter human induced pluripotent stem cell (iPSC) and neural stem cell (NSC) lines. PLoS One 10:e0116032
Merling, Randall K; Sweeney, Colin L; Chu, Jessica et al. (2015) An AAVS1-targeted minigene platform for correction of iPSCs from all five types of chronic granulomatous disease. Mol Ther 23:147-57
Efthymiou, Anastasia G; Chen, Guibin; Rao, Mahendra et al. (2014) Self-renewal and cell lineage differentiation strategies in human embryonic stem cells and induced pluripotent stem cells. Expert Opin Biol Ther 14:1333-44

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