IMBA Stem Cell Core Facility (ISCCF)
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Building on its existing strengths in stem cell research, IMBA has embarked on an ambitious plan to create 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 main focus of the ISCCF is collection of human embryonic stem cells (hESCs) and disease-specific human induced pluripotent stem cells (hiPSCs). Together these will provide significant opportunities to researchers studying disease mechanisms, developing new treatments, and testing the efficacy and safety of new drugs.
The ISCCF also provides a resource of homozygous targeted mouse ES cell lines – a service known as “Haplobank” – to researchers. This valuable resource makes it possible to combine the power of a haploid genome with pluripotency of embryonic stem cells to uncover fundamental developmental and biological processes in defined cell types at the genomic level.
The ISCCF incorporates the existing workflows for cell engineering, including the CRISPR-Cas9 and AAVS2 technologies, to offer state-of-the-art services to researchers.
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, four 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.
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 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 for expansion of hES cell lines, genetically modified and/or non-modified, provided by faculty members and to subsequently freeze and archive them as master cell banks. Lines currently in our repository includeH9, H7 and HES3.
Custom primary cells production
ISCCF offers services on isolation and cryopreservation of primary cells from healthy and patient’s 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 10ml blood
Derivation of fibroblasts from punch skin biopsies
- For this we require a 2-3mm 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 process and time-consuming process, especially when multiple lines are grown at the same time. ISCCF provides services for cell reprogramming and characterization of normal and disease-specific hiPSCs in order to avoid wasteful duplication of efforts across different laboratories on site, across Austria and in the broader scientific community.
We currently offer two 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
- For fibroblasts reprogramming, we require 1 vial of 5 hundred thousand cells.
- For blood reprogramming, we require 2 million cells
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
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Pictures of iPSC colonies were taken from the whole well using the Celigo cytometer under bright field, blue, red and green channels
Multiplex-FISH, mouse haploid ES cells
Custom Genome Engineering
The ISCCF incorporates the existing workflows for cell engineering, including the CRISPR-Cas9 and AAVS2 technologies, to offer state-of-the-art services to researchers. We believe that these valuable resources will enable mechanistic study into modelling many human diseases in an appropriate cell type derived from an individual with the disease.
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, Haplobank generated a genome-wide library of murine embryonic stem cell lines with defined genetic mutations, which 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.
The Haplobank website for more information and ordering of cell lines will be available soon.