how important of Serum proportion of culture medium in cell culture - SOS (Mar/26/2007 )
hi everyone
In our lab, we bought new cell line of L132 epithelial cells. The recommended culture medium by company ATCC is MEM medium with 10% FBS. But culture medium of our old L132 cell line is MEM medium with 5% FBS. Now I don't know for this new cell source, I should strictly respect the recommendation of company (10% FBS) or it will be also okay if I follow the old habit as in our lab (5% FBS). Some elder colleagues told me that our old cells also been recommended culture in 10% FBS medium, but they found cell grow well in 5% FBS medium too.
Acturally I did the test to culture this new cell in both 10% FBS and 5% FBS medium already; my observation is cell growth seems the samely well in both medium. But I am not sure that in the long run, cell growth will be really similar in 10% FBS medium as in 5% FBS medium ? I don't know how important role of Serum proportion of culture medium played in cell culture; there exist some range that cell grow similarly well or this is really a very sensitive parameter in cell culture life?
wait for comment
thanks
-lilylille-
QUOTE (lilylille @ Mar 26 2007, 01:06 AM)
hi everyone
In our lab, we bought new cell line of L132 epithelial cells. The recommended culture medium by company ATCC is MEM medium with 10% FBS. But culture medium of our old L132 cell line is MEM medium with 5% FBS. Now I don't know for this new cell source, I should strictly respect the recommendation of company (10% FBS) or it will be also okay if I follow the old habit as in our lab (5% FBS). Some elder colleagues told me that our old cells also been recommended culture in 10% FBS medium, but they found cell grow well in 5% FBS medium too.
Acturally I did the test to culture this new cell in both 10% FBS and 5% FBS medium already; my observation is cell growth seems the samely well in both medium. But I am not sure that in the long run, cell growth will be really similar in 10% FBS medium as in 5% FBS medium ? I don't know how important role of Serum proportion of culture medium played in cell culture; there exist some range that cell grow similarly well or this is really a very sensitive parameter in cell culture life?
wait for comment
thanks
In our lab, we bought new cell line of L132 epithelial cells. The recommended culture medium by company ATCC is MEM medium with 10% FBS. But culture medium of our old L132 cell line is MEM medium with 5% FBS. Now I don't know for this new cell source, I should strictly respect the recommendation of company (10% FBS) or it will be also okay if I follow the old habit as in our lab (5% FBS). Some elder colleagues told me that our old cells also been recommended culture in 10% FBS medium, but they found cell grow well in 5% FBS medium too.
Acturally I did the test to culture this new cell in both 10% FBS and 5% FBS medium already; my observation is cell growth seems the samely well in both medium. But I am not sure that in the long run, cell growth will be really similar in 10% FBS medium as in 5% FBS medium ? I don't know how important role of Serum proportion of culture medium played in cell culture; there exist some range that cell grow similarly well or this is really a very sensitive parameter in cell culture life?
wait for comment
thanks
Well, serum is critical component of cell culture media because of unique coctail of cytokines and growth factors, which regulate cell growth and differentiation. As I known this cell line is derived from embryonic lung tissue and HeLa positive. May be ATCC recommended FBS % is optimal from terms of survival epithelial cell phenotype. Playing with FBS% may change phenotype of your cell ( note that these cell from embryonic tissue). May be some changes with expression of ceratin will occur. Is it important for you, it depends on your exp aims. I think it will be useful for you to culture this cells in recommended serum % and in parallel with your %. The changes you could see by checking phenotype ( ELISA , FACS against specific antigens) and may be morphology during a long time period. May be it is possible that Hela cells will start up ( a little possibility but why not)?
-circlepoint-
QUOTE (lilylille @ Mar 26 2007, 02:06 AM)
hi everyone
In our lab, we bought new cell line of L132 epithelial cells. The recommended culture medium by company ATCC is MEM medium with 10% FBS. But culture medium of our old L132 cell line is MEM medium with 5% FBS. Now I don't know for this new cell source, I should strictly respect the recommendation of company (10% FBS) or it will be also okay if I follow the old habit as in our lab (5% FBS). Some elder colleagues told me that our old cells also been recommended culture in 10% FBS medium, but they found cell grow well in 5% FBS medium too.
Acturally I did the test to culture this new cell in both 10% FBS and 5% FBS medium already; my observation is cell growth seems the samely well in both medium. But I am not sure that in the long run, cell growth will be really similar in 10% FBS medium as in 5% FBS medium ? I don't know how important role of Serum proportion of culture medium played in cell culture; there exist some range that cell grow similarly well or this is really a very sensitive parameter in cell culture life?
wait for comment
thanks
In our lab, we bought new cell line of L132 epithelial cells. The recommended culture medium by company ATCC is MEM medium with 10% FBS. But culture medium of our old L132 cell line is MEM medium with 5% FBS. Now I don't know for this new cell source, I should strictly respect the recommendation of company (10% FBS) or it will be also okay if I follow the old habit as in our lab (5% FBS). Some elder colleagues told me that our old cells also been recommended culture in 10% FBS medium, but they found cell grow well in 5% FBS medium too.
Acturally I did the test to culture this new cell in both 10% FBS and 5% FBS medium already; my observation is cell growth seems the samely well in both medium. But I am not sure that in the long run, cell growth will be really similar in 10% FBS medium as in 5% FBS medium ? I don't know how important role of Serum proportion of culture medium played in cell culture; there exist some range that cell grow similarly well or this is really a very sensitive parameter in cell culture life?
wait for comment
thanks
Source/quality of serum is very important and ignorance is common place.
In order of price and quality:
British<European<South American<North America<Australian<New Zealand.
For consistancy of experimental results, serum should be BATCH TESTED. Below is what we look for with our serum:-
Cell Viability
Cell Growth
Cell Respiration
iNOS Induction (enzyme)
cGMP Content of cells
Sodium Channel expression
Cytochrome C content
Mitochondrial Electron Transport chain (oxidation/reduction status).
We only use New Zealand serum and even from that region there can be massive variability. We only batch test every 18 months to 2 years. Once tested we buy 60-100 x 500ml bottles and store it.
To answer your question directly, always use the media that is recommended by ATCC/ECACC.
-Rhombus-
QUOTE (Rhombus @ Mar 26 2007, 02:03 AM)
QUOTE (lilylille @ Mar 26 2007, 02:06 AM)
hi everyone
In our lab, we bought new cell line of L132 epithelial cells. The recommended culture medium by company ATCC is MEM medium with 10% FBS. But culture medium of our old L132 cell line is MEM medium with 5% FBS. Now I don't know for this new cell source, I should strictly respect the recommendation of company (10% FBS) or it will be also okay if I follow the old habit as in our lab (5% FBS). Some elder colleagues told me that our old cells also been recommended culture in 10% FBS medium, but they found cell grow well in 5% FBS medium too.
Acturally I did the test to culture this new cell in both 10% FBS and 5% FBS medium already; my observation is cell growth seems the samely well in both medium. But I am not sure that in the long run, cell growth will be really similar in 10% FBS medium as in 5% FBS medium ? I don't know how important role of Serum proportion of culture medium played in cell culture; there exist some range that cell grow similarly well or this is really a very sensitive parameter in cell culture life?
wait for comment
thanks
In our lab, we bought new cell line of L132 epithelial cells. The recommended culture medium by company ATCC is MEM medium with 10% FBS. But culture medium of our old L132 cell line is MEM medium with 5% FBS. Now I don't know for this new cell source, I should strictly respect the recommendation of company (10% FBS) or it will be also okay if I follow the old habit as in our lab (5% FBS). Some elder colleagues told me that our old cells also been recommended culture in 10% FBS medium, but they found cell grow well in 5% FBS medium too.
Acturally I did the test to culture this new cell in both 10% FBS and 5% FBS medium already; my observation is cell growth seems the samely well in both medium. But I am not sure that in the long run, cell growth will be really similar in 10% FBS medium as in 5% FBS medium ? I don't know how important role of Serum proportion of culture medium played in cell culture; there exist some range that cell grow similarly well or this is really a very sensitive parameter in cell culture life?
wait for comment
thanks
Source/quality of serum is very important and ignorance is common place.
In order of price and quality:
British<European<South American<North America<Australian<New Zealand.
For consistancy of experimental results, serum should be BATCH TESTED. Below is what we look for with our serum:-
Cell Viability
Cell Growth
Cell Respiration
iNOS Induction (enzyme)
cGMP Content of cells
Sodium Channel expression
Cytochrome C content
Mitochondrial Electron Transport chain (oxidation/reduction status).
We only use New Zealand serum and even from that region there can be massive variability. We only batch test every 18 months to 2 years. Once tested we buy 60-100 x 500ml bottles and store it.
To answer your question directly, always use the media that is recommended by ATCC/ECACC.
Thank you Rhombus for usefull info about Serum, is these points generally concern also to choosing serum for hybridoma cell culturing or may be any specific suggestions?
-circlepoint-
thanks a lot to Rhombus and circlepoint; I think I have enough reason to respect the ATCC recommenation when culture this cell. also thanx a lot for sharing the common information about serum, which
serves to me very usefully.
-lilylille-
Dear Circlepoint,
Hybridoma cell lines used for production of monoclonals for the clinic are now routinely grown in "chemically defined and protein free media". These products have come along way from when I first tried to do serum free experiments in the late 1970's. Their advantage is that the lack of proteins greatly reduces downstream purification and antigenic problems. New Zealand serum is the highest quality serum and is still the only BSE free region in the world that produces FCS/FBS. As this is clearly the case all our groups cell culture work is done with NZ serum. As stated before it gives higher levels of induced enzymes, higher transfection rates and channel expression in our hands.
-Rhombus-
QUOTE (Rhombus @ Mar 27 2007, 03:39 AM)
Dear Circlepoint,
Hybridoma cell lines used for production of monoclonals for the clinic are now routinely grown in "chemically defined and protein free media". These products have come along way from when I first tried to do serum free experiments in the late 1970's. Their advantage is that the lack of proteins greatly reduces downstream purification and antigenic problems. New Zealand serum is the highest quality serum and is still the only BSE free region in the world that produces FCS/FBS. As this is clearly the case all our groups cell culture work is done with NZ serum. As stated before it gives higher levels of induced enzymes, higher transfection rates and channel expression in our hands.
Hybridoma cell lines used for production of monoclonals for the clinic are now routinely grown in "chemically defined and protein free media". These products have come along way from when I first tried to do serum free experiments in the late 1970's. Their advantage is that the lack of proteins greatly reduces downstream purification and antigenic problems. New Zealand serum is the highest quality serum and is still the only BSE free region in the world that produces FCS/FBS. As this is clearly the case all our groups cell culture work is done with NZ serum. As stated before it gives higher levels of induced enzymes, higher transfection rates and channel expression in our hands.
Thank you Rhombus for detail explanation!
Could you give me advice about Hollow Fiber system efficacy to culture hybridoma cells and Mab production. We will start to use it in our lab now. As I've read one of the main advantages is low compartment for cell growing, allowing to obtain high concentration of Mabs ( no need to concentrate before purification). Another one is circulating oxygen enrichment media that allows to remove toxic metabolite and one more( I think very important but may have deep stones) is that cell growth compartment separates from media delivery system with dialysis membrane with cut off 30 or 50 kDa , so you can use usual serum ( non protein free) and so cytokines can permeable to hybridoma cells but bovine IgG and BSA not, so we are free from the main impurities bIgG and balast protein BSA. It look like invitingly but may be some underlying potential problems occur?. I think also that BSA is important for cell growth as it deliver fatty acids and facilitate other active protiens into the cell , so removal of BSA is not a good idea , is'nt it?
Thank you in advance!
-circlepoint-
QUOTE (circlepoint @ Mar 28 2007, 03:48 AM)
QUOTE (Rhombus @ Mar 27 2007, 03:39 AM)
Dear Circlepoint,
Hybridoma cell lines used for production of monoclonals for the clinic are now routinely grown in "chemically defined and protein free media". These products have come along way from when I first tried to do serum free experiments in the late 1970's. Their advantage is that the lack of proteins greatly reduces downstream purification and antigenic problems. New Zealand serum is the highest quality serum and is still the only BSE free region in the world that produces FCS/FBS. As this is clearly the case all our groups cell culture work is done with NZ serum. As stated before it gives higher levels of induced enzymes, higher transfection rates and channel expression in our hands.
Hybridoma cell lines used for production of monoclonals for the clinic are now routinely grown in "chemically defined and protein free media". These products have come along way from when I first tried to do serum free experiments in the late 1970's. Their advantage is that the lack of proteins greatly reduces downstream purification and antigenic problems. New Zealand serum is the highest quality serum and is still the only BSE free region in the world that produces FCS/FBS. As this is clearly the case all our groups cell culture work is done with NZ serum. As stated before it gives higher levels of induced enzymes, higher transfection rates and channel expression in our hands.
Thank you Rhombus for detail explanation!
Could you give me advice about Hollow Fiber system efficacy to culture hybridoma cells and Mab production. We will start to use it in our lab now. As I've read one of the main advantages is low compartment for cell growing, allowing to obtain high concentration of Mabs ( no need to concentrate before purification). Another one is circulating oxygen enrichment media that allows to remove toxic metabolite and one more( I think very important but may have deep stones) is that cell growth compartment separates from media delivery system with dialysis membrane with cut off 30 or 50 kDa , so you can use usual serum ( non protein free) and so cytokines can permeable to hybridoma cells but bovine IgG and BSA not, so we are free from the main impurities bIgG and balast protein BSA. It look like invitingly but may be some underlying potential problems occur?. I think also that BSA is important for cell growth as it deliver fatty acids and facilitate other active protiens into the cell , so removal of BSA is not a good idea , is'nt it?
Thank you in advance!
Sorry can not help you with either hollow fibre or BSA question. Maybe somebody else out there knows...ask the questions as a new topic?
Rhombus
-Rhombus-
QUOTE (Rhombus @ Mar 28 2007, 05:41 AM)
QUOTE (circlepoint @ Mar 28 2007, 03:48 AM)
QUOTE (Rhombus @ Mar 27 2007, 03:39 AM)
Dear Circlepoint,
Hybridoma cell lines used for production of monoclonals for the clinic are now routinely grown in "chemically defined and protein free media". These products have come along way from when I first tried to do serum free experiments in the late 1970's. Their advantage is that the lack of proteins greatly reduces downstream purification and antigenic problems. New Zealand serum is the highest quality serum and is still the only BSE free region in the world that produces FCS/FBS. As this is clearly the case all our groups cell culture work is done with NZ serum. As stated before it gives higher levels of induced enzymes, higher transfection rates and channel expression in our hands.
Hybridoma cell lines used for production of monoclonals for the clinic are now routinely grown in "chemically defined and protein free media". These products have come along way from when I first tried to do serum free experiments in the late 1970's. Their advantage is that the lack of proteins greatly reduces downstream purification and antigenic problems. New Zealand serum is the highest quality serum and is still the only BSE free region in the world that produces FCS/FBS. As this is clearly the case all our groups cell culture work is done with NZ serum. As stated before it gives higher levels of induced enzymes, higher transfection rates and channel expression in our hands.
Thank you Rhombus for detail explanation!
Could you give me advice about Hollow Fiber system efficacy to culture hybridoma cells and Mab production. We will start to use it in our lab now. As I've read one of the main advantages is low compartment for cell growing, allowing to obtain high concentration of Mabs ( no need to concentrate before purification). Another one is circulating oxygen enrichment media that allows to remove toxic metabolite and one more( I think very important but may have deep stones) is that cell growth compartment separates from media delivery system with dialysis membrane with cut off 30 or 50 kDa , so you can use usual serum ( non protein free) and so cytokines can permeable to hybridoma cells but bovine IgG and BSA not, so we are free from the main impurities bIgG and balast protein BSA. It look like invitingly but may be some underlying potential problems occur?. I think also that BSA is important for cell growth as it deliver fatty acids and facilitate other active protiens into the cell , so removal of BSA is not a good idea , is'nt it?
Thank you in advance!
Sorry can not help you with either hollow fibre or BSA question. Maybe somebody else out there knows...ask the questions as a new topic?
Rhombus
Thank you for reply! I will do so
-circlepoint-