Stem Cell Core Facility

We perform genome-editing in ESc/IPSc in Human and Mouse species.

TYPE OF EDITING

• Knock-outs
• Knock-Ins
• AAVS1/ROSA26

Targeting Approaches

• Electroporation (AMAXA 4D system)
• Lipofection

CRISPR delivery can be DNA or RNP based.

Time: Turnaround time depends on project complexity, but typically it ranges between 8-14 weeks.

Screening and final edited products: We do not perform genotyping on picked clones due to capacity limitations. Instead, internal costumers are tasked for this step. Once successful editing has been identified, the facility will proceed to expand 3-5 edited clones and hand them over as frozen vials. These clones will be also banked in our facility.

REQUEST A GENOME TARGETING SERVICE

To submit a request for Genome Targeting in JIRA: https://jira.imp.ac.at/servicedesk/customer/portal/4; click on Gene Targeting.
If you wish to provide us with your own stem cell line and targeting reagents please attach relevant handling and QC information in the JIRA submission form. Provision of good quality starting material is solely the responsibility of the costumer.  

TYPE OF REPROGRAMMING TECHNOLOGY AND BIOMATERIAL USED

We offer IMBA groups Sendai Virus non-integrating reprogramming technologies under feeder-free conditions to reprogram from healthy and diseased source material:

• Blood (Peripheral Blood Mononuclear Cells (PBMCs).
• Skin biopsies (fibroblasts)

Turnaround time ranges between 12-16 weeks, depending on complexity and disease background status. All lines are regularly checked for mycoplasma.

Upon request we can perform QC services as listed under ‘QUALITY CONTROL SERVICE OF HUMAN STEM CELLS.’

Please note, that the facility must reply with existing ethics laws. It is the customer’s responsibility to provide the relevant patient consent and ethics approval.

REQUEST A HUMAN IPSC REPROGRAMMING SERVICE

To submit a request for Genome Targeting in JIRA: https://jira.imp.ac.at/servicedesk/customer/portal/4; click on ‘Human Reprogramming’.

The IMBA iPSC Biobank, is a collection of induced pluripotent stem cell lines (iPSC lines) produced from skin (fibroblasts) and blood cells (PBMCs). The iPSC Biobank is integrated into the IMBA Stem Cell Core Facility. It stores and provides iPSC lines from clinically normal test subjects as a resource for scientific research.

The iPSC Biobank lines are highly characterized and authenticated using immunological, phenotypic, genotypic, and sterility-based analyses. SCCF scientists perform immunocytochemistry and quantitative analyses to verify the presence of the appropriate markers of pluripotency. To confirm the pluripotent status, the colony morphology and karyotype of the iPSCs are examined. Finally, the cells are authenticated by short tandem repeat (STR) and tested for microbial contamination as well as for the presence of human pathogens.

The resource is available to internal IMBA groups, academic groups, and commercial customers. The ethical guidelines of the iPSC Biobank are always based on prevailing laws and internal regulations. Click the link for more information about the new webshop. https://lnkd.in/ePrUh2Eh

Prolonged culture conditions and genetic manipulation of cells may expose cells to selection pressures and subsequently compromise cell fitness and stem cell properties. We recommend for researchers to test the quality of their lines routinely during their work.

Following QC and reagent services are available for internal IMBA groups:

• SNP array-based genotyping  
• Pluripotency tests: Alkaline Phosphatase (AP) staining pluripotency test, Expression of markers of the undifferentiated state (IF or Flow cytometry analyses), Tri-lineage differentiation
• Cell line submission and banking
• Viral panel testing (CMV, EBV, HBV, HCV, HSV-1, HSV-2, HIV, VZV, HPV) is available from the MBS Facility.
• Home-made stem cell media (E8)

REQUEST A SERVICE IN QUALITY CONTROL IN HUMAN STEM CELLS:

To submit a request in JIRA: https://jira.imp.ac.at/servicedesk/customer/portal/4; click on ‘Quality Control’.

We offer hands-on training to researchers interested to learn stem cell handling.

Find out more on our next HANDS-ON HUMAN STEM CELL WORKSHOP here.