Cell Culture Core
The Cell Culture Core offers cell culture services to investigators on an individual contractual basis. This will be an agreement between Core personnel and the researcher. At that time a plan will be created, with lead time and estimated cost for services.
As part of our quality control program, we have Mycoplasma and Quarantine facilities. The Core will test for and eradicate mycoplasma from cell cultures. All cell culture is performed in a sterile environment employing aseptic techniques.
The Core provides cell banking, with the capacity to freeze and store 10,000 vials. Any cells received for culturing will be quarantined until tested and found to be free from mycoplasma.
The Core has the following equipment:
- Walk-in warm room
- Walk-in cold room
- Spinner apparatus to accommodate 20+L of cells in suspension
- Roller bottle machine to accommodate the culture of 200+ roller bottles
- Mycoplasma and Quarantine facilities
- Automated countess cell counter
- ELISA reader
Hands-on Cell Culture training and lab maintenance and cleaning demonstrations are available regularly. Call 216.444 .5814 to schedule an appointment.
Cell Culture Training
The Core provides training on good lab practices to new researchers. This training can be tailored to individual needs and includes aseptic technique and culturing and maintaining cell lines.
The Core can accommodate the storage of cryovials. This service is offered to Lerner Research Institute researchers for backup storage of precious cell lines.
Direct Mycoplasma Testing
This method uses an enriched agar to support the growth of colonies. Cells and supernatant are swabbed onto agar and incubated in a modular incubator chamber. Samples are viewed microscopically every other day for twoweeks. Mycoplasma contamination is detected by the appearance of a fried egg -like growth.
EBV Transformations/Separation of Whole Blood
The Core can provide immortalization of lymphocytes with the Epstein Barr Virus (EBV). The Core will separate the whole blood, collect the lymphocytes, infect with EBV, and establish a cell line.
Eradication of Mycoplasma from Cell Cultures
The Core will test for and eradicate mycoplasma from cells. If mycoplasma contamination is present in cultures, the Core will begin to eradicate using specialized antibiotics. This will take several weeks to complete. At the end of the treatment, the Core will provide clean cells growing in culture as well as several frozen vials.
Indirect Mycoplasma Testing
The Core also uses a quick method to detect mycoplasma. This kit detects the four most common types of mycoplasma to contaminate cells. This is useful in determining the type of mycoplasma present.
Insect Cell Culture
The Core is equipped with a 27°C incubator that is an optimum temperature to support the growth of insect cells.
The Core routinely performs a direct and an indirect method of testing for mycoplasma. The following two tests are done in parallel.
The cells should be grown without antibiotics for three to four days. Collect cells for testing by scraping the adherent cells and collecting about 5mls of cells and media. For suspension cells, grow without antibiotics and supply 5mls for testing.
Roller Bottle Cultures
The Core has the capability of producing large scale adherent cells cultured in either 850 cm or 1750 cm roller bottles grown on a roller apparatus in a 37°C warm room. This is a very suitable method when large amounts of cells are required. This method also works well with suspension cells.
The Core screens FBS from different sources for optimum cell growth. The batch that proves most compatible to a broad range of cell types is provided for Lerner Research Institute researchers.
Spinner Cultures Cells
Useful for the production of large volumes of suspension cells. Vessels come in various sizes that are used in conjunction with a magnetic stirrer spinner base. The core can provide volumes from 100mls-10L.
The Core offers sterility services using in-house prepared broths along with regular mycoplasma and endotoxin testing. Quarterly testing results are available.
Equipment8'x10' Warm Room
The Cell Culture Core is equipped with a 37°C 8'x10' walk-in warm room which can accommodate large-scale roller bottle cultures. The Core can grow cells in either 850cm2 or 1760cm2 roller bottles. Both suspension and monolayer cells can be grown successfully in this type of room.
Countess® Automated Cell Counter
The Countess® Automated Cell Counter from Invitrogen offers easy and accurate cell counting and viability counts. The counter uses trypan blue staining combined with a sophisticated image analysis algorithm to produce accurate cell and viability counts in just thirty seconds.
The Multiskan FC from Thermo Scientific is a high-quality filter-based microplate photometer. It is used to measure absorbance from appropriate 96 or 384-well plate formats in the wavelength range of 340 to 850nm.
Roller Bottle Machine
A roller bottle machine able to accommodate the cultures of 200+ roller bottles.
Proper Use of Fyrite CO2 Meter (View PDF)
Frequently Asked Questions
What is the cryopreservation capacity in the Cell Culture lab?
Currently we can store 10,000 vials.
How should I prepare my samples for mycoplasma testing?
This information can be found on our iLab webpage.
Should I trypsinize the cells for mycoplasma testing?
No. Scrape cells and supply 5mL in a tube.
I'm new to cell culture, where do I find information on tissue culture techniques?
The Cell Culture Core has several protocols on cell culture:
- Aseptic techniques
- Recovery of cells from liquid nitrogen
- Culturing cells-monolayer and suspension
- Performing a viable count
- Storage of cells in liquid nitrogen
- Quality control
- Specialized techniques
Is there a facility for large-scale cell culture?
The Core has the capacity to grow large volumes of cells, both monolayer, suspension and insect cells.
In addition to the cell services listed, will the Core provide other "special cell cultures" not listed?
Yes, the Core will accommodate any special orders. Researchers should discuss this in advance with Core personnel.
How does the cost for cell services compare with other institutes and outside companies?
Our charges are competitively priced. We charge less than other institutes for certain services and we are much cheaper than outside companies offering the same service.
Why should I purchase cells from the Cell Culture Core if I can get them free from my colleagues?
Cells obtained from outside sources may not always be as healthy as you think. All cells from the Cell Services Core are stored at early passages and are regularly tested for mycoplasma and endotoxins.
The Cell Services Core provides services related to cell culture, preparation of cell media and solutions, and glassware washing and sterilization. The facility strives to provide services that are efficient, convenient, and cost effective to investigators. The core can provide researchers with large quantities of monolayer and suspension cells, and has facilities for specialized cell culture such as spinner and roller bottle culture, insect cell culture, and EBV transformations. The core also has a mycoplasma and quarantine facility and offers mycoplasma testing on a regular basis. Cell culture training can be arranged for new employees.
The Media production lab is a convenient and economical service with outstanding quality control. The core prepares all media in house and can also accommodate custom orders. A wide variety of bacteriological prepared media is available also.
The Glasswash facility handles washing and sterilization of all types of glassware for all labs in the LRI. The facility maintains a variety of sterile laboratory glassware and pipettes in a common storage area. Glassware cabinets are replenished on all floors daily. Custom autoclaving needs can be easily accommodated.