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Stem Cell Core Facility

About

Building on its existing strengths in stem cell research, the IMBA has created a Stem Cell Core Facility (ISCCF) as a part of its initiative for an Austrian Centre for Stem Cell Research. The facility utilizes state of the art technologies for banking and provision of high quality and well-characterized stem cell products.

The goal of the ISCCF is to facilitate stem cell research by providing cutting-edge stem cell technologies and services that are beyond the capacity of individual research groups. ISCCF services are available to researchers on site, across Austria and to the broader scientific community. 

The focus of the ISCCF is the derivation and collection of human embryonic stem cells (hESCs) and disease-specific human induced pluripotent stem cells (hiPSCs). Together, these provide researchers with significant opportunities to study disease mechanisms, develop new treatments, and test the efficacy and safety of new drugs.

The ISCCF also provides a resource of homozygous targeted mouse ESC (mESC) lines – known as “Haplobank” – to researchers (see below).  This valuable resource combines the power of a haploid genome with the pluripotency of embryonic stem cells, making it possible to uncover fundamental developmental and biological processes in defined cell types at the genomic level. 

All the procedures performed at the ISCCF follow Standard Operating Procedures derived from the best practices of biomanufacturing processes. These include sample collection, expansion, and characterization of cell lines which are essential in preparation for banking and distribution. The facility also implements systems for traceability of materials, reagents and products.

The facility has a lab for molecular biology/biochemistry and four separate tissue culture labs: one for quarantine/primary cell culture, one for virus/reprogramming, one for human iPSC/ESC expansion and banking, one for mouse haploid embryonic stem cells and gene targeting.

Services

The ISCCF ensures that all human material from donors is obtained with appropriate ethical approval and that all regulatory requirements are met. This includes informed donor consent and traceability. We also ensure that the agreements that are in place impose no restrictions, other than ethical considerations, on how the samples and accompanying data generated during the project can be used.

Custom hES cell lines expansion and banking

The facility offers a service to expand, freeze and archive hES cell lines, genetically modified and/or non-modified, provided by faculty members. Lines currently in our repository include H9, and H7.

 

Custom primary cell production

ISCCF can isolate and cryopreserve primary cells from healthy and patient donors such as:

  • Derivation of peripheral blood mononuclear cell (PBMC) from whole blood:
    • For this we require 10 ml whole blood (Normal or disease cohort)
    • We typically cryopreserve 6 to 10 million PMBCs per 10 ml blood
  • Derivation of fibroblasts from punch skin biopsies
    • For this we require a 2-3 mm skin punch biopsy
    • We cryopreserve 4 vials (2 million cells per vial)
    • Turnaround time:  About 25 days

Custom hiPSC Generation:

The generation, maintenance and differentiation of hiPSCs is a labor-intensive and time-consuming process, especially when multiple lines are grown simultaneously. ISCCF provides services for cell reprogramming and characterization of normal and disease-specific hiPSCs, avoiding wasteful duplication of efforts across different laboratories on site, across Austria and in the broader scientific community.

We currently offer non-integrating reprogramming technologies:

  • Sendai Virus system
    • Turnaround time: 3-4 months
  • Episomal, Epstein-Barr Nuclear Antigen 1 plasmid system
    • Turnaround time: About 3-4 months

Please note:

  • For fibroblast reprogramming, we require 1 vial of 500,000 cells.
  • For blood reprogramming, we require 2,000,000 cells.

 We offer the following quality control tests & characterization:

  • Pluripotency and the capacity to differentiate into the three germ layers: we perform differentiation in vitro using lineage specific markers (n=2 per germ layer) and quantify them using an automated imaging instrument
  • Genetic stability testing by array-based genotyping. For these assays, samples of the original tissue will be used as controls
  • Cell line identity check will be performed using short tandem repeat profiling to assure correct identity of iPSC lines and to rule out any cross-contamination
  • Mycoplasma testing will be performed prior to cryopreservation

Custom Genome Engineering

ISCCF incorporates the existing workflows for cell engineering, including the CRISPR genome editing in human ES/iPSC and mouse ES cell gene targeting technologies, to offer state-of-the-art services to researchers. These valuable resources will enable mechanistic studies and modelling of many human diseases, in an appropriate cell type derived from an individual with the disease.

Custom Quality Control tests & Characterization

We offer the following quality control tests & characterization:

  • Pluripotency and the capacity to differentiate into the three germ layers. We perform differentiation in vitro using lineage specific markers (n=2 per germ layer) and quantify them by an automated imaging instrument
  • Genetic stability testing by array-based genotyping. For these assays, samples of the original tissue will be used as controls
  • Cell line identity check will be performed using short tandem repeat profiling to assure correct identity of iPSC lines and to rule out any cross-contamination
  • Karyotyping by Multiplex Fluorescence In Situ Hybridization (M-FISH)
  • Mycoplasma testing will be performed prior to cryopreservation

Research & Development

ISCCF will fundamentally embrace the need for method optimization and robustness of hPSCs bioprocesses. Dedicated R&D work within the facility includes:

  • Improving existing processes and implementing the latest technologies to develop robust, cost-effective, and scalable hPSCs bioprocesses.
  • Developing a Good Manufacturing Protocol-compliant PSC expansion process.
  • Networking with other groups, notably with renowned academic partners, for formal transfer of methodology into the facility and conversion into an operational process.  This will ensure that the method is performed as intended within the originated laboratories environment.
  • Ensuring that the facility operates at the highest level of international bench-marking for the hPSCs bioprocesses.

HAPLOBANK

In the post-genomic era, science is accumulating large datasets from high-throughput screening. Validation of hits from such screens still requires genetic testing. To this end, we created Haplobank, a genome-wide library of murine ESC lines with defined genetic mutations. Haplobank contains over 100,000 conditionally mutated and reversible mESC clones, targeting about 70% of the protein-coding genome (almost 17,000 genes). The resource is available to all scientists, and represents the largest ever library of homozyogous mutant embryonic stem cell lines to date. Haplobank overcomes issues arising from clonal variability, because mutations can be repaired in single cells and at whole genome scale. In addition, the haploid genome of these embryonic stem cells enables reverse genetics. Haplobank can be used to functionally validate predictions or to study in-depth phenotypic effects of gene loss, thereby helping to phenotypically annotate the mouse genome.

For more information about the Haplobank and ordering of cell lines visit the website (www.haplobank.at)

Team

AGU Chukwuma Allison

AGU Chukwuma Allison

Head of SCCF

+43 1 79044 4880

BUCHMANN Arabella

BUCHMANN Arabella

Research Technician

+43 1 79044 4878

DOBLINGER Max

DOBLINGER Max

Research Technician

+43 1 79044 4879

FARINA LOPEZ Sara

FARINA LOPEZ Sara

Research Assistant

+43 1 79044 4270

MIREA Madalina Alexandra

MIREA Madalina Alexandra

Research Assistant

+43 1 79044 4558

VOLZ Carol Jennifer

VOLZ Carol Jennifer

Staff Scientist

+43 1 79044 4886

FOONG-SOBIS Michelle

FOONG-SOBIS Michelle

Technical Assistant

+43 1 79044 4723

KINZER Manuela

KINZER Manuela

Research Assistant

+43 1 79044 4884

SCHULLER Nicole

SCHULLER Nicole

Research Assistant

+43 1 79044 4885

STEMBERGER Roman Ferdinand

STEMBERGER Roman Ferdinand

Technical Assistant

+43 1 79044 4303

VAN DE LOO Nina

VAN DE LOO Nina

Research Assistant

+43 1 79044 4890

WETZEL Ellen

WETZEL Ellen

Research Assistant

+43 1 79044 4882