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How much PBS can be added to the culture medium?
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Adrian Wiechoczek
How much PBS can be added to the culture medium?
Hello Chaoqun Huang,
Its seems that the drug of your interest is water soluble. If it is so, then I would suggest to make the stock solution fist in sterile distilled water and then final working concentrations in Dulbecco′s Modified Eagle′s Medium (DMEM) culture medium. Make sure that the solvent (here water or PBS) final amount should not literally exceed more than 1% in culture medium in order to avoid disturbance in physical, chemical proprieties, and concentrations of other component of the culture medium.
DMEM is a modification of Basal Medium Eagle (BME) that contains four-fold concentrations of the amino acids and vitamins. The original formulation contained 1000 mg/L of glucose and was used to culture embryonic mouse cells. Since then, it has been modified in several ways to support primary cultures of mouse and chicken cells, as well as a variety of normal and transformed cells. It is a widely used basal medium for supporting the growth of many different mammalian cells includingprimary fibroblasts, neurons, glial cells, smooth muscle cells, as well as cell lines such as HeLa, 293, Cos-7, and PC-12 etc.
Hence, addition of large amount of solvent (here water or PBS) may alter/dilute the constituents of DMEM specifically made for specific type of cell culture or purpose. In any condition, try to ensure that the solvent present in final working concentration is not more that 1% (v/v) in DMEM culture medium in order to retain the osmolarity and other ingredients concentration in culture medium.
Its seems that the drug of your interest is water soluble. If it is so, then I would suggest to make the stock solution fist in sterile distilled water and then final working concentrations in Dulbecco′s Modified Eagle′s Medium (DMEM) culture medium. Make sure that the solvent (here water or PBS) final amount should not literally exceed more than 1% in culture medium in order to avoid disturbance in physical, chemical proprieties, and concentrations of other component of the culture medium.
DMEM is a modification of Basal Medium Eagle (BME) that contains four-fold concentrations of the amino acids and vitamins. The original formulation contained 1000 mg/L of glucose and was used to culture embryonic mouse cells. Since then, it has been modified in several ways to support primary cultures of mouse and chicken cells, as well as a variety of normal and transformed cells. It is a widely used basal medium for supporting the growth of many different mammalian cells includingprimary fibroblasts, neurons, glial cells, smooth muscle cells, as well as cell lines such as HeLa, 293, Cos-7, and PC-12 etc.
Hence, addition of large amount of solvent (here water or PBS) may alter/dilute the constituents of DMEM specifically made for specific type of cell culture or purpose. In any condition, try to ensure that the solvent present in final working concentration is not more that 1% (v/v) in DMEM culture medium in order to retain the osmolarity and other ingredients concentration in culture medium.
From an osmotic point of view, it's not a problem, that's why we are using PBS and not water.
However, you'll dilute nutrients and serum components (growth factors etc) and you're affecting the buffer system, which usually is relying on bicarbonate, HEPES, or alike.
Have a look at the errors/variations you'll have from batch to batch when preparing media. So a variation of 0.1 to 0.5% (i.e 1 to 5 ml per litre) should not pose a problem. In any case, you'll have a PBS only internal control that is lacking your substance of interest.
From an osmotic point of view, it's not a problem, that's why we are using PBS and not water.
However, you'll dilute nutrients and serum components (growth factors etc) and you're affecting the buffer system, which usually is relying on bicarbonate, HEPES, or alike.
Have a look at the errors/variations you'll have from batch to batch when preparing media. So a variation of 0.1 to 0.5% (i.e 1 to 5 ml per litre) should not pose a problem. In any case, you'll have a PBS only internal control that is lacking your substance of interest.
Would tend to add things at a low concentration ie 0.1 to 1 % or so at a general rule.Of course if things are critical, test the effect of eg addition of pbs , in advance to confirm suitabity.
Would tend to add things at a low concentration ie 0.1 to 1 % or so at a general rule.Of course if things are critical, test the effect of eg addition of pbs , in advance to confirm suitabity.
Usually a few percent is OK. The answer depends on many variables, especially your cell line, medium, and culture routine. Most importantly, as your experiment compares the effect of medium with and without added drug, you must use the results from diluted medium for both drug and control.
Usually a few percent is OK. The answer depends on many variables, especially your cell line, medium, and culture routine. Most importantly, as your experiment compares the effect of medium with and without added drug, you must use the results from diluted medium for both drug and control.
Hello Chaoqun Huang,
Its seems that the drug of your interest is water soluble. If it is so, then I would suggest to make the stock solution fist in sterile distilled water and then final working concentrations in Dulbecco′s Modified Eagle′s Medium (DMEM) culture medium. Make sure that the solvent (here water or PBS) final amount should not literally exceed more than 1% in culture medium in order to avoid disturbance in physical, chemical proprieties, and concentrations of other component of the culture medium.
DMEM is a modification of Basal Medium Eagle (BME) that contains four-fold concentrations of the amino acids and vitamins. The original formulation contained 1000 mg/L of glucose and was used to culture embryonic mouse cells. Since then, it has been modified in several ways to support primary cultures of mouse and chicken cells, as well as a variety of normal and transformed cells. It is a widely used basal medium for supporting the growth of many different mammalian cells including primary fibroblasts, neurons, glial cells, smooth muscle cells, as well as cell lines such as HeLa, 293, Cos-7, and PC-12 etc.
Hence, addition of large amount of solvent (here water or PBS) may alter/dilute the constituents of DMEM specifically made for specific type of cell culture or purpose. In any condition, try to ensure that the solvent present in final working concentration is not more that 1% (v/v) in DMEM culture medium in order to retain the osmolarity and other ingredients concentration in culture medium.
Good luck
Hello Chaoqun Huang,
Its seems that the drug of your interest is water soluble. If it is so, then I would suggest to make the stock solution fist in sterile distilled water and then final working concentrations in Dulbecco′s Modified Eagle′s Medium (DMEM) culture medium. Make sure that the solvent (here water or PBS) final amount should not literally exceed more than 1% in culture medium in order to avoid disturbance in physical, chemical proprieties, and concentrations of other component of the culture medium.
DMEM is a modification of Basal Medium Eagle (BME) that contains four-fold concentrations of the amino acids and vitamins. The original formulation contained 1000 mg/L of glucose and was used to culture embryonic mouse cells. Since then, it has been modified in several ways to support primary cultures of mouse and chicken cells, as well as a variety of normal and transformed cells. It is a widely used basal medium for supporting the growth of many different mammalian cells including primary fibroblasts, neurons, glial cells, smooth muscle cells, as well as cell lines such as HeLa, 293, Cos-7, and PC-12 etc.
Hence, addition of large amount of solvent (here water or PBS) may alter/dilute the constituents of DMEM specifically made for specific type of cell culture or purpose. In any condition, try to ensure that the solvent present in final working concentration is not more that 1% (v/v) in DMEM culture medium in order to retain the osmolarity and other ingredients concentration in culture medium.
Good luck
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From an osmotic point of view, it's not a problem, that's why we are using PBS and not water.
However, you'll dilute nutrients and serum components (growth factors etc) and you're affecting the buffer system, which usually is relying on bicarbonate, HEPES, or alike.
Have a look at the errors/variations you'll have from batch to batch when preparing media. So a variation of 0.1 to 0.5% (i.e 1 to 5 ml per litre) should not pose a problem. In any case, you'll have a PBS only internal control that is lacking your substance of interest.
From an osmotic point of view, it's not a problem, that's why we are using PBS and not water.
However, you'll dilute nutrients and serum components (growth factors etc) and you're affecting the buffer system, which usually is relying on bicarbonate, HEPES, or alike.
Have a look at the errors/variations you'll have from batch to batch when preparing media. So a variation of 0.1 to 0.5% (i.e 1 to 5 ml per litre) should not pose a problem. In any case, you'll have a PBS only internal control that is lacking your substance of interest.
More
VOTE
Would tend to add things at a low concentration ie 0.1 to 1 % or so at a general rule.Of course if things are critical, test the effect of eg addition of pbs , in advance to confirm suitabity.
Would tend to add things at a low concentration ie 0.1 to 1 % or so at a general rule.Of course if things are critical, test the effect of eg addition of pbs , in advance to confirm suitabity.
More
VOTE
Usually a few percent is OK. The answer depends on many variables, especially your cell line, medium, and culture routine.
Most importantly, as your experiment compares the effect of medium with and without added drug, you must use the results from diluted medium for both drug and control.
Usually a few percent is OK. The answer depends on many variables, especially your cell line, medium, and culture routine.
Most importantly, as your experiment compares the effect of medium with and without added drug, you must use the results from diluted medium for both drug and control.
More
VOTE