SC3:Single Cell and Cell Culture Facilities
- The current cell and tissue culture facilities are organized in four rooms situated on two floors. 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 for analyses at the single cell level, we have recently acquired a FACS Melody (BD Biosciences). As well, small sample technologies have been developped : RTqPCR on small samples using preamplification steps (with Fluidigm Biomark or classical qPCR instruments), transgenic methods such as TRAP (translating ribosome affinity purification). Both transcriptomic and genomic (DNA methylation, histone marks analyses) are and will be performed.
Equipment and expertise :
a) Cell/tissue culture rooms (L. Blanchon, Rachel Guiton):
- 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 (S. Maupetit, N. Allègre):
This is a virtual facility, using different instruments that are available at the GReD and on the Clermont-Fd campus:
- 1 FACS Melody (BD Biosciences), that can sort single cells per well
- 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)