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Keeping Cells Happy: Topics in Cell Health Maintenance and Viability

Sep 13, 2018 at 12:00 PM ET

Abstract

Good practices are vital for safeguarding the health of cells in long-term culture. Cell health, maintenance, and viability cover a wide range of topics, from ensuring that cells are healthy in culture to having confidence that the cells are characterized correctly. This presentation will focus on topics such as cell viability assays that measure cell growth, authentication of cell lines through short tandem repeat (STR) profiling, mycoplasma testing, aseptic technique, and cryopreservation. From services to techniques, we will discuss ATCC’s role in ensuring the health and maintenance of cells in culture.

Key Points

  • Mycoplasma infection can chronically affect the well-being of cells in culture without causing obvious symptoms. Screening is required to determine if cell cultures are infected.
  • STR profiling detects misidentification or cross-contamination of human cell lines.
  • When culturing specialty cells, such as stem cells or primary cells, certain considerations regarding the choice of media and reagents must be taken.
  • Proliferation kits and reagents such as XTT, MTT, and Reliablue™ are great tools for measuring cell health and growth.

Presenter

Steve Budd, headshot.

Steven Budd, MS, MBA

Product Line Business Specialist, ATCC

Steven Budd is a Product Line Business Specialist that manages the cell culture reagents at ATCC. He has 4 years of experience in the product management of scientific tools. Before that, he gained 4 years of experience in biomedical research and cell culture as a research specialist at the University of North Carolina at Chapel Hill. Mr. Budd has a M.S. in Biology from the University of North Carolina at Wilmington and an M.B.A. from North Carolina State University.

Questions and Answers

Can you keep the freezing container, the coolcell, for long periods, like weeks?

You should not.  It is designed to freeze cells slowly at -70°C or -80°C for a 4 hours to overnight. After that, they need to go into the -140°C, this is the only way to stop metabolic action, anything longer at -70°C and you could risk cell damage.

Cell culture media can be able with or without phenol red? Any advantages or disadvantages?

Phenol red in of itself does not really make much of a difference in the livelihood of cells. Whether it’s there or not, the cells will grow.  But without phenol red, it will just be clear, so there will be no visible indicator of pH.  One thing to note, mentioned in the presentation, is that phenol red can behave like a hormone in that it may bind to estrogen receptors on mammary cells.  So it could read as a false positive.  

Does the mycoplasma detection kit quantify or identify the mycoplasma contaminant?

Neither to both.  It is designed to be sensitive enough to read to femto levels of DNA, so it should indicate any presence of mycoplasma, and only mycoplasma. No other bacteria or microbe will be read.  Its PCR based, so the reading is either a yes or no as to whether or not there is contamination. It can’t identify the exact species, because it was that sensitive, it would miss any other species. So it’s better to just let you know if you have contamination and deal with the culture accordingly. Which should be to get rid of it.

If a cell culture is contaminated with fungus or bacteria, what can be done to rescue the cells?

The only advice I can give is to toss the contaminated cell cultures. If you try to rid the media of contamination, you probably will not be successful, and if it’s in an incubator with other cells, you run the risk of contaminating other cell cultures.

What is the difference between Stem Cell qualified FBS and Standard FBS?

No real difference between the two.  We test certain lots with stem cells and test for growth. If it passes the growth test to satisfaction, we reserve that lot with the Stem cell qualified part number.