Cell Biology
The Cell Biology platform meets the needs of the Institute’s research teams for all methodological aspects applicable to the cell biology of human pluripotent cells and their derivatives. It also ensures the quality control of these cells and, if necessary, the optimization of their culture protocols.
Quality control is a crucial and determining factor for the appropriate use of human pluripotent stem cells and their derivatives. At I-Stem, we are particularly vigilant in this field of investigation because undetected cellular anomalies can lead to major errors in pathological modeling and pharmacology, or introduce severe risk factors in the context of cell therapy. This is why our team is in charge of the systematic and regular verification of all the cells generated in the Institute.
We use karyotyping to detect chromosomal abnormalities ranging in size from 5 to 10Mb (Megabases). This allows us to highlight structural abnormalities (reciprocal translocation), as well as number abnormalities (trisomy, monosomy). We use classical cytogenetic techniques such as G or R-banding but also the multicolor Fluorescence In Situ Hybridization (mFISH) technique.
The team has a complete cytogenetic equipment including a CDS-5 (Thermotron) drying chamber and a Metasystem automated microscopy station. This allows to obtain very good quality karyotypes for an optimal classification of chromosomes and the precise visualization of complex chromosome breakage and rearrangement points.
We also regularly perform genome analysis of cells using SNP microarrays (Illumina) marketed by the company Integragen (Evry, France). We can thus localize the exact size (from 100 Kilobases) of genomic anomalies detected in human pluripotent cells and their derivatives.
Finally, to explore the genomic integrity of the biological material of cells, our platform regularly tests new detection technologies. Our collaboration with the company StemGenomics (Montpellier, France) has allowed us to propose a rapid detection test for the most recurrent anomalies acquired by pluripotent cells. This test allows the very fast verification of our cells at key and critical steps of the bioprocess, such as the creation of large cell banks by the Bioproduction Platform or the editing of the cell genome by the CRISPR/Cas9 method.
The team has acquired a real expertise in these fields over the last few years. Current projects include the exploration of immune bypass pathways (“ghost cells”) with Thierry Heidmann’s team, the company Viroxis and the CR2TI of Nantes or projects in the framework of the MyoPharm program, in which I-Stem collaborates with other French research teams on ultra-rare neuromuscular diseases.
We also offer karyotyping services (by G or R-banding but also by mFISH) for external academic or industrial teams.
ANR AAPG 2021 – PRCE: Ghosting The Cells (GTC) :
Project to create “ghost” cells that maintain the HLA antigen presentation systems intact but induce immunotolerance by presenting an immunosuppressive molecule from a retrovirus at the membrane.
Team members
Karine Giraud-Triboult
Platform Manager (CECS)
Karine is a Functional Genomics Engineer. She joined the team at I-Stem’s creation in January 2005, working on the DM1 project. She has headed the Cell Biology Platform since 2017.
Lina El Kassar
Platform Engineer (CECS)
Lina has a PhD in Biology. She joined the team at I-Stem’s creation in January 2005, working on the DM1 project. She led the iPS workshop for three years, and since 2016 has been in charge of monitoring the genomic integrity of cells at I-Stem.
Louise Penez
Platform Engineer (INSERM)
Louise joined the team at the beginning of 2022 as an M2 trainee. After graduating with a degree in Biotechnology, she joined the team as a research engineer. She is working on the Ghosting The Cells pre-clinical program.
Hamel Mahiou
Technician (CECS)
Hamel joined the team at the end of 2022 as a work-study student. After obtaining her professional degree in biotechnology, she joined the team as a platform technician. She provides support for all team and laboratory projects, particularly in cell culture and quality control.
Collaborations
Specialist in the biology of infectious and endogenous retroviruses, and more particularly in the molecular basis of their induction of immunosuppression.
Specialist in immunology at VIROXIS which develops immunosuppressive molecules derived from retroviral proteins for transplant applications.
Scientific leader of the humanized rodents platform of LabEx IGO, which focuses on the development of strategies to control the immune response.
– INSERM, Centre de Recherche en Myologie, Sorbonne Université, UMRS974, INSERM, Institut de Myologie-Faculté de Médecine de la Pitié Salpêtrière, Antoine Muchir
– CNRS UMR 5310 – INSERM U1217, Institut NeuroMyoGène, Université de Lyon, Arnaud Jacquier, Laurent Shaeffer
– INSERM U955 IMRB UPEC, Créteil, Frederic Relaix
– INSERM U1251, Marseille Medical Genetics, Valérie Delague
Publications
Generation of heterozygous and homozygous NF1 lines from human-induced pluripotent stem cells using CRISPR/Cas9 to investigate bone defects associated with neurofibromatosis type 1.
01 January 2024
Frontiers in cell and developmental biology
Establishment of heterozygous and homozygous SHANK3 knockout clonal pluripotent stem cells from the parental hESC line SA001 using CRISPR/Cas9.
01 October 2023
Stem cell research
CRISPR/Cas9-mediated generation of human embryonic stem cell sub-lines with HPRT1 gene knockout to model Lesch Nyhan disease.
01 September 2023
Stem cell research
Thermotron
Thermostatically controlled drying chamber CDS-5
Metasystem
Automated microscopy station. The exact location of chromosome breakpoints, karyotypes and chromosome classification are obtained with Isis and Ikaros software.
QIACUBE
Automated DNA and RNA extraction using columns.