Stem Cell Facility

The Stem Cell Core Unit supports scientists working with human embryonic stem cells (hESC) and human induced pluripotent stem cells (hiPSC) with the focus on the generation of patient or disease-specific iPSC. The core unit is closely associated to the Institute of Experimental Pharmacology and Toxicology, especially to those groups working on cardiac tissue engineering and interested in pharmacological studies of cardiovascular disease using hiPSC.

Recently, it was shown that adult somatic cells (e.g. fibroblasts) can be reprogrammed to iPSC via a defined combination of transcription factors [1,2]. Since this groundbreaking discovery, the underlying methods were further developed and a lot of information has been published [3-5]. At the moment, the generation of patient- or disease-specific iPSC seems to be especially helpful to study disease mechanisms and offers new possibilities for the development and study of novel drugs or therapies [6].

Services

human fibroblasts
Human fibroblast cultur derived from dermal tissue
embryoid bodies
"embryoid bodies" (EBs) from hiPS cells in suspension cultur
iPS
hiPS cell cultur on feeder-cells
hiPSC clone
hiPSC clone: immunofluorescence staining Oct4=red, TRA-1-60=green

The facility routinely keeps cultures of human pluripotent stem cells and continuously reprograms human adult fibroblasts into hiPSC. To this end, different methods of reprogramming (integrating and non-integrating) as well as several options for characterisation of pluripotency have been established. Currently, genome editing in hiPSC via the CRISPR/Cas9 system is analysed which would allow for insertion, removal or replacement of DNA sequences in the genome.

The following services are offered by the facility. However, at the beginning of each project a detailed plan should be worked out individually.

  • Individual counseling for optimal project realization


  • training of hESC / hiPSC handling and methods
  • generation of patient-specific iPSC
    • Establish primary cell culture from patient sample (mostly fibroblasts from skin biopsies; alternatively blood-derived cells) à estimated duration ca. 3 weeks
    • Reprogramming is routinely accomplished using non-integrating viral vectors (Sendai virus, CytoTune kit) for the delivery of the reprogramming factors à estimated duration ca. 3 weeks
    • Selection, expansion and cryo-conservation of newly produced hiPS clones à estimated duration ca. 2 months
  • characterization of hiPSC
    • morphology
    • expression profile à detection of stem cell and pluripotency markers via RT-qPCR and immunohistochemistry or immunofluorescence
    • potential for in vitro differentiation à „embryoid body formation“ à confirmation of development of all 3 germ layers via RT qPCR
  • quality control of ES / iPS cell cultures
  • creation of a patient-specific hiPS bank
  • supply with conditioned medium (for hESC or hiPSC in feeder-free culture)
  • supply with feeder cells (mouse embryonic fibroblasts (MEF))
  • counselling and support for the development of protocols for differentiation into different cell types
  • counselling and support for genome editing of hiPSC using the CRISPR/Cas9 system

Registration

Please contact for further information by phone or email to the contact address listed below and note the agreement of the Facility.

Contact

References

[1] Takahashi, K., et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131, 861-72 (2007)
[2] Yu, J., et al. Induced pluripotent stem cell lines derived from human somatic cells. Science 318, 1917-20 (2007)
[3] Nakagawa, M., et al. Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nature Biotechnol. 26, 101-6 (2008)
[4] Lin, T., et al. A chemical platform for improved induction of human iPSCs. Nature Meth. 6 (11), 805-9 (2009)
[5] Warren, L., et al. Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. Cell Stem Cell 7, 618-630 (2010)
[6] Park, I.-H., et al. Disease-specific induced pluripotent stem cells. Cell 134, 877-886 (2008)