The CBM has 6 biosafety level 2 laboratories for the use of cell cultures, with a total of 91 booths, distributed on floors 1, 2, 3, 4 and -1, with the same characteristics on floors 1 to 4. On the 1st floor there is a P2 laboratory for microbiology and on floor -1 the P2 laboratory is dedicated to work with viruses.

In addition, the CBM has a P3 laboratory that is used exclusively for experiments involving human and/or animal cell cultures, human and/or animal viruses, and parasites requiring biosafety level 3.

In these laboratories the service staff is responsible for:

Management of purchase of products necessary for the preparation of media for cell culture.
Management of the purchase of the necessary equipment.
Preparation and sterilization of media and solutions used for the different work in cultures.
Control of the proper functioning and preventive maintenance of the equipment in the general culture room (incubators, cabinets, pipettors, etc.).
Control of the general cleanliness of the laboratories.
General maintenance of the laboratories (cleaning of equipment, removal of plastic and glass waste, replacement of material).
Elaboration of cleaning and maintenance protocols.
Elaboration of rules of use of the facility.
Control of invoicing per user.
Detection of mycoplasmas in the cultures of users who request this service.

Mycoplasma detection is carried out in cell lines and culture media by means of a colorimetric assay, based on the ability to recognize “TOLL-LIKE-RECEPTOR 2” mycoplasmas and to induce the activation of NF-kB and other transcription factors. In the presence of mycoplasmas TLR2 expressed on the surface of HEK-blue cells, activates these transcription factors which, in turn, induce the secretion of alkaline phosphatase, a protein easily detectable by the blue staining of the medium.

The assay will be performed periodically, informing users at least 5 days in advance, so that they can collect the samples to be tested and leave them in the place marked for this purpose by the service.

For the elimination of mycoplasmas in contaminated cultures, the service offers antibiotic aliquots and the protocol to follow.

General use material in the different laboratories and services of the CBM

The Material Washing Service provides support to the Research Groups, as well as to other Services of the Center that require it.

The staff of the Service is in charge of the collection, cleaning and sterilization of the laboratory material and its orderly storage, leaving it available for collection by the users.

They are also in charge of the sterilization of all the media, buffers, solutions etc. Both those prepared in the Service and those prepared in the laboratories and require sterilization.

Location: Floor 2, Laboratory 210

Material for use in P2 culture laboratories

The personnel of the Service are in charge of the collection of material from the culture laboratories and the deactivation of different residues, as well as the cleaning and sterilization of the laboratory material and its orderly storage, leaving it available for collection by the users.
Verification of the operation of the autoclaves by means of periodic controls (Vacuum Test, Bowie & Dick Test, and Biological Test).

The media laboratory, where different media, buffer solutions, etc. are prepared, offers the user a permanent stock, with the possibility of placing orders.

Preparation of media and solutions for cells of animal origin

A permanent stock of the following media and solutions is offered:
MEM 1X
RPMI
RPMI + hepes
PBS 1X
PBS 10X
PBS 1X compl.
BSS
DMEM 1X
DMEM 2X
TC-100
Antimycotic
Glutamine
EDTA
Gentamicin
M. antibiotics
AANE 100X
Trypsin EDTA
Sodium pyruvate
Trypsin
Special media
Agar1.4% Agar
Agar 1%
Neutral red 1,4+ Agar
M 3
A.C.D
DEAE-dextran
lysis buffer
HBSS
These media and solutions are prepared under maximum safety conditions, being carried out in a sterile laminar flow cabinet and filtered with 0.22 µm membranes. Subsequently, a sterility test is performed.
Location: Floor 2, Laboratory 211 and Floor -1, Laboratory 032.

Preparation of culture media for microorganisms

Liquid media are prepared following the corresponding protocols according to demand and sterilized by heat in a 121ºC liquid cycle for 20 minutes.
Solid media. Prepared with agar and heat sterilized in autoclave in liquid cycle 121ºC, 20 minutes.

A permanent stock of the following media and solutions is available:
LB
LB+agar
Plates with LB+agar medium
Plates with LB+agar+antibiotic medium
2x YT medium
Location: Floor 1, Lab 123

Preparation of solutions and buffers

They are prepared following the corresponding protocol (Molecular Cloning appendix, etc.) with controls on pH and physical characteristics. They are mostly sterilized by steam autoclaving or by filtration with a 0.22 µm membrane in a sterile cabinet.

A permanent stock of the following media and solutions is available:
ECL (developer solution)
TBE 10X
TRIS GLYCINE 10X
TBS 10X
PBS 10X
TAE 50X
Mg Cl2 1M
K Ac 5M
SSC 20X
TRIS 1.5M pH 8.8
TRIS 1M pH 6.8
TRIS 1M pH 7.5
TRIS 1M pH 8
TRIS 1M pH 8.8
SDS 10% SDS
SDS 20% SDS
EDTA 0.5 M pH 8
Na Cl 5M
Na Ac 3M pH 5.2
Monosodium Phosphate 1M
Disodium Phosphate 1M
RIPA
PB 0,2 N Ph:7,4
Location: Floor 2, Laboratory 211
Serum treatment

Filtration by 0.22 μm, in sterile laminar flow cabinet.
Sterility test.

Preparation of sustenance medium for Drosophila flies

It is performed with different components, using Agar as solid support. Subsequently, it is sterilized and dispensed in tubes, bottles, etc.
Location: Floor 4, Laboratory 413

The Media Preparation Service is associated with the Sterilization Service and has all the necessary equipment (balances, pH meters, shakers, refrigerators, etc.) for the preparation and storage of culture media (e.g., LB, LB-agar) and sterile solutions (e.g., PBS, EDTA, TBE, TAE, Tris-HCl pH 7.4, etc.) that are in common use and are available to all groups in the Center.

To request any of the services that are not in stock we have prepared a form (download here) to be sent as an attachment to the address peticion-medios@cbm.csic.es, filled in with your data. In the “Request” box there is a drop-down box where you can mark the type of medium or solution you are requesting (TC SOLUTIONS, MICROBIOLOGY MEDIA, JR CELL MEDIA, P2 VIRUS MEDIA). The quantity and description of what is requested will be indicated below. Requests will be processed and two days later you will be able to pick up your order.

Only requests received in this way will be attended. This guarantees that your requests will be carried out by one of the technicians, even if some of the staff are on leave.