Gibco Serum-Free Media (SFM)

Gibco serum-free media promotes superior cell growth and viability

We are the world’s largest manufacturer of serum-free cell culture media and a leader in the development of innovative specialty media, including serum-free media options. Serum-free media (SFM) allow researchers to grow a specific cell type or perform a specific application in the absence of serum.

Advantages of using serum-free media include:

  • Easier purification and downstream processing
  • Precise evaluations of cellular function
  • Increased definition
  • More consistent performance
  • Increased growth and/or productivity
  • Better control over physiological responsiveness
  • Enhanced detection of cellular mediators

How to convert from serum-supplemented to serum-free media

When you're ready to switch to serum-free culture, we're ready to help. Whichever adaptation method you choose (sequential or conditioned medium), we strongly recommend that you always take these precautions:

  • Make a frozen stock of the cells in the serum-supplemented medium prior to adaptation
  • As a precaution, maintain a culture of the cells in each prior condition when starting the next level of adaptation in case the cells do not survive in the next passage

Sequential adaptation

Sequential adaptation is our preferred method for adapting cells to serum-free media (SFM), with a typical conversion being:

Passage 1 75% serum-supplemented medium:25% SFM
Passage 2 50% serum-supplemented medium:50% SFM
Passage 3 25% serum-supplemented medium:75% SFM
Passage 4 100% SFM

Because the change from 75% to 100% SFM may be too stressful for your cells, you may need to carry the cells for 2–3 passages in a 10% serum-supplemented medium: 90% SFM mixture. Most cell lines can be considered fully adapted after 3 passages in 100% SFM. Occasionally you may have trouble getting your cells past a certain step even before going 100% SFM. If this happens, go back and passage the cells 2–3 times in the previous ratio of serum-supplemented media to serum-free media.

Adaptation with conditioned medium

An alternate method for adaptation to SFM involves the use of conditioned medium. This is medium the cells have been growing in for one full passage. If you choose this method, you can facilitate adaptation as follows:

Passage 1 100% serum-supplemented medium
Passage 2 50% medium from passage 1:50% SFM
Passage 3 50% medium from passage 2:50% SFM
Passage 4 50% medium from passage 3:50% SFM
Passage 5 100% SFM

Points to consider in serum-free culture

Overall, cells in serum-free culture are more sensitive to extremes of pH, temperature, osmolality, mechanical forces, and enzyme treatment.

Antibiotics

It is best not to use antibiotics in serum-free media. If you do, we recommend that you use 5- to 10-fold less than you would in a serum-supplemented medium. This is because serum proteins tend to bind a certain amount of the antibiotic added; without these serum proteins the level of antibiotic may be toxic to certain cells.

Higher density

Cells must be in the mid-logarithmic phase of growth with viability >90% prior to adaptation. Sequential adaptation may be necessary.

Seeding cultures at a higher density than normal at each passage during SFM adaptation may help the process. Because some percentage of cells may not survive in the new culture environment, having more cells present will increase the number of viable cells to further passage.

Clumping

Cell clumping often occurs during adaptation to SFM. We recommend that you gently triturate the clumps to break them up when passaging cells.

Morphology

It is not uncommon to see slight changes in cellular morphology during and after adaptation to SFM. As long as doubling times and viability remain good, slight changes in morphology should not be a reason for concern.

Sequential adaptation

Sequential adaptation is our preferred method for adapting cells to serum-free media (SFM), with a typical conversion being:

Passage 1 75% serum-supplemented medium:25% SFM
Passage 2 50% serum-supplemented medium:50% SFM
Passage 3 25% serum-supplemented medium:75% SFM
Passage 4 100% SFM

Because the change from 75% to 100% SFM may be too stressful for your cells, you may need to carry the cells for 2–3 passages in a 10% serum-supplemented medium: 90% SFM mixture. Most cell lines can be considered fully adapted after 3 passages in 100% SFM. Occasionally you may have trouble getting your cells past a certain step even before going 100% SFM. If this happens, go back and passage the cells 2–3 times in the previous ratio of serum-supplemented media to serum-free media.

Adaptation with conditioned medium

An alternate method for adaptation to SFM involves the use of conditioned medium. This is medium the cells have been growing in for one full passage. If you choose this method, you can facilitate adaptation as follows:

Passage 1 100% serum-supplemented medium
Passage 2 50% medium from passage 1:50% SFM
Passage 3 50% medium from passage 2:50% SFM
Passage 4 50% medium from passage 3:50% SFM
Passage 5 100% SFM

Points to consider in serum-free culture

Overall, cells in serum-free culture are more sensitive to extremes of pH, temperature, osmolality, mechanical forces, and enzyme treatment.

Antibiotics

It is best not to use antibiotics in serum-free media. If you do, we recommend that you use 5- to 10-fold less than you would in a serum-supplemented medium. This is because serum proteins tend to bind a certain amount of the antibiotic added; without these serum proteins the level of antibiotic may be toxic to certain cells.

Higher density

Cells must be in the mid-logarithmic phase of growth with viability >90% prior to adaptation. Sequential adaptation may be necessary.

Seeding cultures at a higher density than normal at each passage during SFM adaptation may help the process. Because some percentage of cells may not survive in the new culture environment, having more cells present will increase the number of viable cells to further passage.

Clumping

Cell clumping often occurs during adaptation to SFM. We recommend that you gently triturate the clumps to break them up when passaging cells.

Morphology

It is not uncommon to see slight changes in cellular morphology during and after adaptation to SFM. As long as doubling times and viability remain good, slight changes in morphology should not be a reason for concern.

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