Current Cell and Tissue Culture Facilities
- The current cell and tissue culture facilities are present on both sites and are organized independently. Different cell lines (transformed or stem cells) and tissues (embryos, primary cultures) from several species (mouse, human, drosophila) are cultured. Classical transgenesis is performed (lipofection, electroporation, lentivirus infection) under the GMO rules (accreditation 6318, April9th 2013 for St Jacques). The authorization to work on human samples is pending.
Due to the need of several teams, single cell/small sample technologies have been developped. Different strategies have been already developped: RTqPCR on small samples (single mouse blastocyst for example) using preamplification steps (with Fluidigm Biomark or classical qPCR instruments), transgenic methods such as TRAP (translating ribosome affinity purification), Flow Analysis Cell Sorting (cell type enrichment from animals tissues or cultured cells), recombinant mix of embryonic cells to regenerate chimeric tissue in vivo. Both transcriptomic and genomic (DNA methylation, histone marks analyses) are and will be performed. Genome editing technologies (CRIsPR/CAS9) are also performed.
Equipment and expertise :
a) Cell/tissue culture rooms:
- 8 Cell culture hoods
- 10 CO2 incubators
- 4 inverted microscopes (with 2 equipped with fluorescence)
- classical material (centrifuge, waterbaths...)
In the future, one of these rooms will be upgraded towards a BSL3.
b) Single cell/small sample equipment:
This is a virtual facility, using different instruments that are available at the GReD and on the Clermont-Fd campus:
- 3 ROCHE Light Cyclers (GReD)
- 1 Fluidigm Biomarck (Gentyane facility, INRA Clermont-Fd) performing single cell qPCR
- 1 ATTUNE Cytometer (Life Technologies)
- 2 micromanipulators for microinjection (Eppendorf) (shared with Mouse Trangenesis Platform Facility - Cézeaux)
- 1 FACS (BD FACSARIA SORP) (CHU Clermont-fd), with 5 lasers, 4 nozzles sizes and with an "autoclone module" that can sort single cells per well