Cell culture and viral vectors

Research support unit

Since 1987, the Cell Culture Unit has been made up of three rooms where cells obtained from multicellular organisms are generated “in vitro”, especially neurons, astrocytes and oligodendrocytes.
obtained from animal brains. The basic functions of the Unit are referred to below:
– Management, cleaning and maintenance of the three rooms.
– Calibration and sterility of incubators.
– Coordination and supervision of the proper use of the equipment.
– Daily sterilization of solid material, and liquid on alternate days.
– Weekly sterilization of thermosensitive material.
– Preparation of reagents according to the needs.
– Testing of fetal sera for batch purchases.
– Filling and maintenance of drums of the cell bank with liquid nitrogen.
– Internal waste management.
– Bi-monthly control and distribution of Service expenses according to the time spent in the room.

Since the year 2000, the possibility of making reagents, media, buffers, agar plates, competent bacteria, etc., has been incorporated into the service, in order to facilitate conventional work for users. According to RD 664/1997, of May 12, on the protection of workers against risks related to exposure to biological agents, the handling of these crops requires 2 different levels of safety (containment level):
– Containment level 1: in the Culture and Established Lines Room. Required for culture of non-primate non-transformed animal cells. With 4 horizontal laminar flow cabins for preferential protection of the samples and another 4 vertical laminar flow cabins that provide protection to the operator as well as to the sample.
– Containment level 2: in Rooms P2-A and P2-B where cell cultures of primates (tumor or non-tumor), human cell lines without intervention, cell cultures of non-primates producing viruses, handling of non-human biological agents are generated. airborne infectious with tropism
human (retrovirus, lentivirus, adenovirus). They have 2 biosafety cabinets that guarantee protection for the operator and also provide protection for the sample.

Equipment

Room P1: Contains 4 horizontal flow cabinets, which allow working with magnifying glasses to perform primary cultures, and 3 vertical flow cabinets, which can be used for both primary cultures and established cell lines. There are also 4 incubators in this room with pre-established temperature and CO2 for cultures of primary origin, and another one for exclusive use for cell lines.

Room P2-B: Contains a type II-A biological safety cabinet, a vertical flow cabinet with 30/70 recirculation, and a vertical flow cabinet without recirculation. The uses assigned to each cabin are indicated in the table below. It has two incubators, one of which is exclusively used for viral vectors.

Room P2-A: Contains a type II-A biological safety cabinet, and an incubator. The use of this room is indicated for the agents that appear in the box below.

Autoclave service

The Cell culture Unit is in charge of the autoclave service. Users must download their duly labeled material before the time established for each service.

– Solid cycle: Monday, Wednesday and Friday at 10:30 a.m.
– Liquid cycle: Monday, Wednesday and Friday at 12 noon.

Staff