The Cell Culture Platform (CCP) was created to cover the needs of the many researchers at the Biodonostia HR, associated centers and external companies working in cell culture to provide them with technical and scientific assistance.
The Platform offers technical assistance and services in the field of cell culture, fluorescence microscopy, flow cytometry and analysis of cellular metabolism (SeaHorse). It also offers cryopreservation services.
The Cell Culture Platform is responsible for the correct operation and preventive maintenance of culture equipment (cabinets, incubators, etc.), the control of the general cleaning of the rooms, managing the appropriate documentation and preparation of cleaning and maintenance protocols, paying particular attention to waste management, biosafety plus microbiological and mycoplasma control.
In 2021, with the acquisition of new infrastructures, the Cell Culture Platform has given shape to two new Units:
- Confocal Microscopy Unit.
- Flow Cytometry Unit.
The Cell Culture Platform of the Biodonostia Health Research Institute forms part of the Spanish Network for Advanced Optical Microscopy (REMOA).
The Cell Culture Platform offers access to infrastructure that entails having prior knowledge of the equipment or, otherwise, undertaking a specific course for this purpose given by the staff. Even more, the Platform offers methodological and scientific support in the design of projects as well as in carrying out budget experiments.
The following services and infrastructure are available at the platform:
Cell culture rooms
LThe Cell Culture Platform has approximately 75 m2 along three cell culture rooms: two of the rooms are reserved to primary and stable cell culture whilst the third one is where virus infections are carried out and it has a biosafety level 2.
All the cell culture rooms are equipped with vacuum and channeling of CO2 y N2 and all have work stations for those gases attached to the incubators to guarantee safety.
Cell Culture room and tool technical information
Los servicios prestados vinculados con las salas de cultivos son las siguientes:
- Cell Collection: human or mouse tissue processing (skin, muscle, etc.) with the purpose of obtaining primary cells (fibroblasts, myoblasts, different types of stem cells, etc.) requested by the researcher.
- Cell culture of stable or immortalized lines: notifying of the commercial existence of the requested cell type, its maintenance, amplification and cryopreservation.
- Cell culture maintenance under strict sterility conditions to assure the correct experiment execution.
- Cell proliferation and viability trials.
- Transfection trials.
- Polluted mycoplasma detection.
- Basic services offered by the Platform: Basic technical teaching and training about handling and experimentation in cell culture, and the maintenance of stocks of cell lines in liquid nitrogen.
Confocal Microscopy Unit
The Cell Culture Platform offers access to the LSM 900 KMAT confocal microscopy with Zeiss AXIO OBSERVER 7. This combines the excellent image quality of the confocal LSM 900 with a Colibri LED fluorescence illumination system with reflection points at 385nm, 475nm, 555nm and 631nm, thereby covering a range of fluorescence from DAPI to Cy5.
The Platform also has a Zeiss Axiovert 200M fluorescence microscope and another Nikon ECLIPSE 80i fluorescence microscope, both with filters for DAPI, FITC, Texas Red, Triple and Far-red for the observation and analysis of very different kinds of samples (cells, tissues, etc.). Similarly, we can observe samples without fluorescence such as haematoxylin-eosin, DAB (3.3’ diaminobenzidine), Jenner-Giemsa, etc.
The Platform has a basic IT system to process and analyze the digital images obtained.
The Platform staff offers immunofluorescence studies plus training and advice to users in the operation of the available methodologies and equipment.
Flow Cytometry Unit
The Platform offers access to the Sony Sorter SH800S which permits the sorting of different cell sizes using the 70 μm, 100 μm and 130 μm microfluidic chips.
It provides a highest level of automation available for any cell size, meaning that researchers can set up, align, calibrate and monitor sorting with the push of a button.
The system software is intuitive and supports sorting in tubes and plates of 96 and 384 wells. The software generates FCS 3.0 and FCS 3.1 files which can be exported tor use with third-party analysis tools.
The Cell Culture Platform also offers the use of the Guava easyCyte 8HT follow cytometer with a blue laser (488 nm) and a red laser (640 nm) which makes possible to view up to 6 fluorescent colors and 2 light scatter. It also enables samples to be processed in one single 96-wll plate.
The Platform also offers advice in flow cytometry analytics basic principles and its application.
Seahorse
The Cell Culture Platform includes the Seahorse XFe96 Analyzer which allows different cellular metabolism measure:
- Mitochondrial function: basal respiration, ATP turnover, proton leak to quantify the cellular bioenergetics; identify mitochondrial functions and measure the cellular response to stress.
- Glycolytic function: it measures parameters such as glycolysis, glycolytic capacity and glycolytic reserve.
- Oxidation of fatty acids.
For further information:
http://www.agilent.com/en-us/promotions/xftechnologyoverview
Liquid nitrogen room
The room has two liquid nitrogen tanks located in the cryogenics room, which are connected to a central alarm system. There is also an oxygen detection alarm in that same room.
The Platform offers cryovials maintenance in liquid nitrogen.
