iPS microscopy

Pluripotent Stem Cells

Sebastian Diecke

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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).

One main objective of the Pluripotent Stem Cell Core (PSCC) at the MDC is to provide scientists at the MDC and Berlin with standardized protocols to derive, maintain and manipulate pluripotent stem cells (PSCs). Additionally the facility offers a reprogramming service and access to characterized induced pluripotent stem cells (iPSCs) as well as to more specialized cell types. In cooperation with the individual researchers the core facility also helps to establish and validate new linage specific differentiation protocols. Another focus of the core is the targeted iPS cell engineering using Crispr/Cas9 or TALEN technology generating reporter as well as knock in or knock out human iPSC cell lines.

We also offer project consulting using iPSCs technologies like directed differentiation and genome engineering and individual hands on training courses.

The PSCC was established September 2014 by funding of the Berlin Institute of health (BIH) and the German Center for Cardiovascular Disease (DZHK) to further increase the basic and translational research activity in Berlin.

So just contact us and we try to help.

Further information can be found on the BIH Stem Cells website.

Pluripotent Stem Cells

Induction of pluripotency is one of the most important scientific discoveries within the last few years. This technique allows researchers to obtain pluripotent stem cells without the controversial destruction of embryos, providing a novel and powerful tool for disease modeling and drug screening approaches.

Induced pluripotent stem cells (iPSCs) have characteristics and differentiation capacities similar to embryonic stem cell and also give rise to tissues of all three germ layers as well as whole organisms. Therefore iPSCs provide an unlimited source of proliferating cells without loss of functionality. The use of iPSCs has tremendous advantages in broad research areas of molecular mechanisms of diseases, drug testing, development of regeneration therapies and may pave the avenue towards personalized medicine.

Discovery

The description of the induction of pluripotency dramatically altered the previous dogma of cellular differentiation as a unidirectional, nonreversible developmental process, resulting in a paradigm shift in the field of developmental biology. For the breakthrough discovery Sir John B. Gurdon and Shinya Yamanaka were jointly awarded with the Nobel Prize in Physiology or Medicine 2012 "for the discovery that mature cells can be reprogrammed to become pluripotent”.

Service and Technology

From somatic cell isolation to application

Somatic cell isolation

Fibroblasts, Peripheral blood mononuclear cells, urinary cells

Fibroblasts

Time from biopsy to start of reprogramming: 4-6 weeks

Peripheral blood mononuclear cells

Time from phlebotomy to start of reprogramming: 6-12 Days

Urinary cells

 

Reprogramming

Integrating viruses, Non-Integrating techniques, Sendai/ Adenovirus

Integrating viruses

Non-Integrating techniques  

Sendai/ Adenovirus 

Characterization of human iPSC

Validation of pluripotency, quality control

Validation of pluripotency

  • Staining for the pluripotency markers (IF + FACS)
  • Differentiation potency (spontaneous/directed)
  • Teratoma
  • Scorecard, Pluritest (planned)

Quality control

  • Vial testing (HIV, HCV, HBV)
  • Mycoplasma + Sterility
  • Genetic integrity/stability
  • Absence of reprogramming virus
  • Identity

 

Banking and  provision

Generation of cell banks, Provision of reference cell lines and differentiated cells

Differentiation

Development of basic differentiation protocols

  • Differentiation of iPSCs into specialized cell types
  • Provision of protocols
  • Support during the establishment of new/published protocols

Standardized protocols

Development and provision of standardized protocols

  • Obtaining and characterization of somatic cells for reprogramming
  • Culture of human pluripotent stem cells (hPSC) in defined conditions
  • Feeder cell co-culture of hPSC
  • Characterization of human pluripotent cells
  • Quality control of hPSC and feeder cultures
  • Mouse and human feeder cells (MEF / HFF)
  • Derivation of human hiPSC
  • Genetic manipulation of hPSC
  • Cryopreservation and Banking of hPSC
  • Differentiation of hPSC

already 52 validated protocols available

Training / Project Consulting

Lectures and hands-on experience

Training is offered on a regular basis:

Basic training

  • Lectures and hands-on experience covering methodology and major techniques used for working with hPSCs
  • focus on maintenance, preservation and characterization

Advanced training

  • Topics will include differentiation, derivation, genetic manipulation

Individual training possibilities

upon request

In collaboration with the AG Kühn improvement or development of new techniques for genome engineering, reprogramming or differentiation.

 

Genome editing

Design and distribution of CRISPR and TALEN constructs, Generation of knockout, reporter and isogenic control cell lines

Transcription activator-like effector nuclease (TALEN) & CRISPR/Cas9 system

  • Design and distribution of CRISPR and TALEN constructs (gene, disruption, gene correction, insertion of reporter constructs)
  • Generation of knockout, reporter and isogenic control cell lines