Core Facility Stem Cells
Pluripotent stem cells are unique tools for closing the gap between the identification of disease-causing molecules or cellular mechanisms and the development of suitable therapeutic approaches. Specifically the development of patient specific disease models will be of major importance.
The mission of the BIH Core Facility Stem Cells is to support basic and translational research by facilitating all aspects of human induced pluripotent stem cell (hiPSC) technology including the derivation, differentiation and distribution of humaniPS cell lines. Additionally, the facility will provide scientists with state of the art protocols and techniques for proper handling and manipulation of human pluripotent stem cells. Furthermore, the Core Facility will organize several hands-on training courses of standardized pluripotent stem cell culture techniques.
The facility has two locations to best serve the researchers at BIH, one at the Charité Campus Virchow-Klinikum (CVK, Wedding), and the other is located at the Max Delbrück Center for Molecular Medicine in the Helmholtz Association (MDC, Berlin-Buch).
Location Charité (CVK))
|Dr. Harald Stachelscheid (Facility leader)||Dr. Sebastian Diecke (Facility leader)|
|Judit Küchler (Technical assistent)||Dr. Jürgen Stumm (Postdoc)|
|Kristin Fischer (Technical assistent)||Narasimha Swami Telugu (research assistent)|
|Janine Cernoch (Technical assistent)||Norman Krüger(Technical assistent)|
|Tanja Fisch (Technical assistent)||Anna Iwanska (Technical assistent)|
|Claudia Schaar (Administration)||Silke Kurths (Technical assistent)|
|Sandra Schommer-Leitner (Technical assistent)|
|Carolin Genehr (Technical assistent)|
|Anna Parsiegel (Apprentice)|
|Prof. Norbert Hübner*: Research group "Genetics and Genomics of Cardiovascular Diseases"||Prof. Georg Duda*: Director Julius Wolff Institute for Biomechanics and Musculoskeletal Regeneration, BCRT|
|Dr. Ralf Kühn: Research group "iPS cell based disease modeling"||Prof. Petra Reinke: Nephrology, Transplantation medicine, BCRT|
|Prof. Ana Pombo: Research group "Epigenetic regulation and chromatin architecture"||Prof. Ulf Landmesser: BIH professorship for cardiology|
|Prof. Thomas Willnow: Research group "Molecular Cardiovascular Research"||Prof. Christian Rosenmund: Neuroscience with the focus on cellulatr and molecular Neurobiology, spokesperson NeuroCure|
|Prof. Clemens Schmitt: Tumor genetics and cellular stress responses|
|* current speaker|
A. Isolation of primary cells form patient samples
e.g. skin, blood, urine
B. Reprogramming of cells into induced pluripotent stem cells
application of non integrating vectors e.g. Sendai Virus, mRNA
C. Characterization and Quality Control of hPSC
- Expression of pluripotency markers (FACS, IF, PCR)
- Whole Genome Gene Expression Profiling (PluriTest)
- Differentiation potential (differentiation into cells of the three germ layers,
- genetic integrity (SNP Array, KaryoBoB)
- Identity (STR Fingerprinting)
- microbial contamination (sterility, mykoplasma, human viral pathoges)
D. Gene editing and cell labeling
Gene KnockOut, KnockIn of transgenes e.g. GFP, introduction and correction of mutations using CRISPR/Cas or TALEN
E. Provision of hiPSC reference lines and banking
- production and validation of cell banks
- the Core Facility established a range of fully characterized hiPSC lines (e.g. BIHi001-A , BIHi004-A) which can be used as a reference
- an overview of the provided cell lines can be found in the human Pluripotent Stem Cell Registry (hPSCreg).
F. Establishment of differentiation protocols and provision of hiPSC derived differentiated cells
e.g. cardiomyocytes, endothelial cells, neurons, neural stem cells, renal progenitor cells, hepatocytes
e.g. cerebral organoids, kidney organoids
H. Provision of infrastructure und equipment
FACS (MACSQuant VYB), RT-PCR (QuantStudio 6), Phase-contrast-, stereo- and fluorescence-microscopes, transfection (Amaxa, Neon), hypoxia-incubators, Picking Hoods
I. Provision of reagents
e.g. special media, viral/non-viral vectors (in cooperation with the Viral Core Facility of the Charité)
J. Provision of standardized protocols
the core provides it users a constantly growing portfolio of state of the art protocols that includes methods for culture, characterization and quality control, cryo-preservation, differentiation and genetic manipulation
K. Project consultancy
support for project planning, grand application and ethical approvals
The generation and differentiation, but also establishment of disease models, require a profound knowledge and hands-on expertise of maintaining and manipulating hiPSCs in tissue culture.
The processes involved in these aspects of hPSC handling in tissue culture are labor intensive and distinct from that of other mammalian cell cultures. The BIH Core Facility Stem Cells organizes regular training on these methodologies.
The training is intended for researchers and technicians who are planning projects using human iPSCs and/or will start working with hiPSCs in the near future.
The next training session will be from 8-10 November 2017.