Since then a number of improved formulations have been used, adding various growth factors or reducing serum content. If cost is your main driver, then the above media is going to be most cost effective and has a long track record. Depending on what you are trying to study, the high serum concentration may swamp whatever you are interested in, so you may end up needing to use a modern, low serum commercial formulation, which will be 2-3x more expensive then the simple one desribed above. Personally, I had very good luck with Lifeline Cell Technology cells and media: http://www.lifelinecelltech.biz/category_s/161.htm Tech support was very good and the sales reps made life very easy. Prices were very good, but I'm not sure about european distribution. You may want to look up EGM or EGM-2 from Lonza which should be more readily available.
If you need to act fast and go with what you got, I'd lean to DMEM 20% FBS plus L-Glut.
Since then a number of improved formulations have been used, adding various growth factors or reducing serum content. If cost is your main driver, then the above media is going to be most cost effective and has a long track record. Depending on what you are trying to study, the high serum concentration may swamp whatever you are interested in, so you may end up needing to use a modern, low serum commercial formulation, which will be 2-3x more expensive then the simple one desribed above. Personally, I had very good luck with Lifeline Cell Technology cells and media: http://www.lifelinecelltech.biz/category_s/161.htm Tech support was very good and the sales reps made life very easy. Prices were very good, but I'm not sure about european distribution. You may want to look up EGM or EGM-2 from Lonza which should be more readily available.
If you need to act fast and go with what you got, I'd lean to DMEM 20% FBS plus L-Glut.
Hello Daniel Moreno Martinez I'd like to tell you about my experience with HUVEC (from ATCC).
As the basal medium, I use F12K nutrient mixture supplemented with 10% FBS, 0.1 mg/mL heparin sodium salt, and 0.03 mg/mL endothelial cell growth supplement (ECGS). This time, I'm using the entire medium by combining all of the elements listed above and changing them every two to three times per week.
This approach, I believe, will be beneficial to you.
Hello Daniel Moreno Martinez I'd like to tell you about my experience with HUVEC (from ATCC).
As the basal medium, I use F12K nutrient mixture supplemented with 10% FBS, 0.1 mg/mL heparin sodium salt, and 0.03 mg/mL endothelial cell growth supplement (ECGS). This time, I'm using the entire medium by combining all of the elements listed above and changing them every two to three times per week.
This approach, I believe, will be beneficial to you.
I can only support the answer from Irfan. With the EGM-2 from Lonza you have the best growth. Although it is more expensive than DMEM or others, you get a more specific endothelial population and you can get 2-3 more population doublings, at the end you can profit with this investment in more and better cells.
I can only support the answer from Irfan. With the EGM-2 from Lonza you have the best growth. Although it is more expensive than DMEM or others, you get a more specific endothelial population and you can get 2-3 more population doublings, at the end you can profit with this investment in more and better cells.
Hi Daniel, I am going to use very similar medium for HUVECs. Could you please let me know, if the protocol is suitable for signalling pathways elucidation by detection of protein phosphorylation (in case you have try it with your cells)? Thank you in advance. Best regards, Stoyan
Hi Daniel, I am going to use very similar medium for HUVECs. Could you please let me know, if the protocol is suitable for signalling pathways elucidation by detection of protein phosphorylation (in case you have try it with your cells)? Thank you in advance. Best regards, Stoyan
Hi Daniel, Could you please give me details about the media you use, i wil use HUVEC, which M199 you use? I found many different formula, it is red phenol free? Or with earl's salt or without, i will order one so, could you please give me cat #? Thanks in advance
Hi Daniel, Could you please give me details about the media you use, i wil use HUVEC, which M199 you use? I found many different formula, it is red phenol free? Or with earl's salt or without, i will order one so, could you please give me cat #? Thanks in advance
You can use DMEM or M199 based medium for only several passages, afterward, the cells would change their morphology and tend to stop growing. By the way, HUVEC is very sensitive to FBS concentration. Too low or too high are both harmful.
You can use DMEM or M199 based medium for only several passages, afterward, the cells would change their morphology and tend to stop growing. By the way, HUVEC is very sensitive to FBS concentration. Too low or too high are both harmful.
Dear Daniel, Based upon our previous work, we found the following formula works very well: - M199 - 20% Fetal bovine serum - Pencilin/Streptomycin 2 mmol/L. - Amphotericin B 2 mmol/L - Glutamine 2mmol/L - HEPES 10 mmol/L - Endothelial cell growth supplement (ECGs) 30-50 µg/ml - Heparin 100 µg/ml Best
Dear Daniel, Based upon our previous work, we found the following formula works very well: - M199 - 20% Fetal bovine serum - Pencilin/Streptomycin 2 mmol/L. - Amphotericin B 2 mmol/L - Glutamine 2mmol/L - HEPES 10 mmol/L - Endothelial cell growth supplement (ECGs) 30-50 µg/ml - Heparin 100 µg/ml Best
This topic has been recently discussed:
https://www.researchgate.net/post/What_is_the_most_cost-effective_media_for_culturing_primary_HUVECs
In general, much of the original work with HUVECs was done in M199 supplemented with 20% FBS and 2 mM glutamine plus penn/strep. This was the original formulation of Jaffe in the seminal paper linked below:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC302542/pdf/jcinvest00186-0087.pdf
Since then a number of improved formulations have been used, adding various growth factors or reducing serum content.
If cost is your main driver, then the above media is going to be most cost effective and has a long track record.
Depending on what you are trying to study, the high serum concentration may swamp whatever you are interested in, so you may end up needing to use a modern, low serum commercial formulation, which will be 2-3x more expensive then the simple one desribed above. Personally, I had very good luck with Lifeline Cell Technology cells and media:
http://www.lifelinecelltech.biz/category_s/161.htm
Tech support was very good and the sales reps made life very easy. Prices were very good, but I'm not sure about european distribution. You may want to look up EGM or EGM-2 from Lonza which should be more readily available.
If you need to act fast and go with what you got, I'd lean to DMEM 20% FBS plus L-Glut.
This topic has been recently discussed:
https://www.researchgate.net/post/What_is_the_most_cost-effective_media_for_culturing_primary_HUVECs
In general, much of the original work with HUVECs was done in M199 supplemented with 20% FBS and 2 mM glutamine plus penn/strep. This was the original formulation of Jaffe in the seminal paper linked below:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC302542/pdf/jcinvest00186-0087.pdf
Since then a number of improved formulations have been used, adding various growth factors or reducing serum content.
If cost is your main driver, then the above media is going to be most cost effective and has a long track record.
Depending on what you are trying to study, the high serum concentration may swamp whatever you are interested in, so you may end up needing to use a modern, low serum commercial formulation, which will be 2-3x more expensive then the simple one desribed above. Personally, I had very good luck with Lifeline Cell Technology cells and media:
http://www.lifelinecelltech.biz/category_s/161.htm
Tech support was very good and the sales reps made life very easy. Prices were very good, but I'm not sure about european distribution. You may want to look up EGM or EGM-2 from Lonza which should be more readily available.
If you need to act fast and go with what you got, I'd lean to DMEM 20% FBS plus L-Glut.
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protocol: Circ Res. 2009;104(5):589-99 Media were purchased by PromoCell, but other brands should also work (e.g. LONZA).
protocol: Circ Res. 2009;104(5):589-99 Media were purchased by PromoCell, but other brands should also work (e.g. LONZA).
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VOTE
For HUVEC growth EGM-2 media with supplement kit + 5%FBS are essential, if You want to harvest it is better to use EBM-2 as working media.
For HUVEC growth EGM-2 media with supplement kit + 5%FBS are essential, if You want to harvest it is better to use EBM-2 as working media.
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Hi
We have been using M199 supplemented with 20% fbs, ecgs and heparin, and it works like a charm!
Hi
We have been using M199 supplemented with 20% fbs, ecgs and heparin, and it works like a charm!
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VOTE
Hello Daniel Moreno Martinez
I'd like to tell you about my experience with HUVEC (from ATCC).
As the basal medium, I use F12K nutrient mixture supplemented with 10% FBS, 0.1 mg/mL heparin sodium salt, and 0.03 mg/mL endothelial cell growth supplement (ECGS). This time, I'm using the entire medium by combining all of the elements listed above and changing them every two to three times per week.
This approach, I believe, will be beneficial to you.
Hello Daniel Moreno Martinez
I'd like to tell you about my experience with HUVEC (from ATCC).
As the basal medium, I use F12K nutrient mixture supplemented with 10% FBS, 0.1 mg/mL heparin sodium salt, and 0.03 mg/mL endothelial cell growth supplement (ECGS). This time, I'm using the entire medium by combining all of the elements listed above and changing them every two to three times per week.
This approach, I believe, will be beneficial to you.
More
VOTE
I can only support the answer from Irfan. With the EGM-2 from Lonza you have the best growth. Although it is more expensive than DMEM or others, you get a more specific endothelial population and you can get 2-3 more population doublings, at the end you can profit with this investment in more and better cells.
I can only support the answer from Irfan. With the EGM-2 from Lonza you have the best growth. Although it is more expensive than DMEM or others, you get a more specific endothelial population and you can get 2-3 more population doublings, at the end you can profit with this investment in more and better cells.
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Hello Daniel, did you had good results using DMEM?
Diogo
Hello Daniel, did you had good results using DMEM?
Diogo
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can you grow HUVEC cells without ECGS in ham's F-12
can you grow HUVEC cells without ECGS in ham's F-12
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Hi Daniel,
I am going to use very similar medium for HUVECs. Could you please let me know, if the protocol is suitable for signalling pathways elucidation by detection of protein phosphorylation (in case you have try it with your cells)? Thank you in advance.
Best regards,
Stoyan
Hi Daniel,
I am going to use very similar medium for HUVECs. Could you please let me know, if the protocol is suitable for signalling pathways elucidation by detection of protein phosphorylation (in case you have try it with your cells)? Thank you in advance.
Best regards,
Stoyan
More
VOTE
Hi Daniel,
Could you please give me details about the media you use, i wil use HUVEC, which M199 you use? I found many different formula, it is red phenol free? Or with earl's salt or without, i will order one so, could you please give me cat #?
Thanks in advance
Hi Daniel,
Could you please give me details about the media you use, i wil use HUVEC, which M199 you use? I found many different formula, it is red phenol free? Or with earl's salt or without, i will order one so, could you please give me cat #?
Thanks in advance
More
VOTE
You can use DMEM or M199 based medium for only several passages, afterward, the cells would change their morphology and tend to stop growing. By the way, HUVEC is very sensitive to FBS concentration. Too low or too high are both harmful.
You can use DMEM or M199 based medium for only several passages, afterward, the cells would change their morphology and tend to stop growing. By the way, HUVEC is very sensitive to FBS concentration. Too low or too high are both harmful.
More
VOTE
Dear Daniel,
Based upon our previous work, we found the following formula works very well:
- M199
- 20% Fetal bovine serum
- Pencilin/Streptomycin 2 mmol/L.
- Amphotericin B 2 mmol/L
- Glutamine 2mmol/L
- HEPES 10 mmol/L
- Endothelial cell growth supplement (ECGs) 30-50 µg/ml
- Heparin 100 µg/ml
Best
Dear Daniel,
Based upon our previous work, we found the following formula works very well:
- M199
- 20% Fetal bovine serum
- Pencilin/Streptomycin 2 mmol/L.
- Amphotericin B 2 mmol/L
- Glutamine 2mmol/L
- HEPES 10 mmol/L
- Endothelial cell growth supplement (ECGs) 30-50 µg/ml
- Heparin 100 µg/ml
Best
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VOTE