I need to start working with this cell line. I´ve found that cells should be seeded at 2-3x10^5 and split when reaching 8x10^5. Other suggest higher densities for seeding, splitting, and freezing. What densities would you recommend for seeding, splitting, and freezing? I will probably work on T25 flasks. Thank you very much!
I need to start working with this cell line. I´ve found that cells should be seeded at 2-3x10^5 and split when reaching 8x10^5. Other suggest higher densities for seeding, splitting, and freezing. What densities would you recommend for seeding, splitting, and freezing? I will probably work on T25 flasks. Thank you very much!
Dear Mª Carmen Ocaña and Denise Grana, I'm having problems with culture of this line at the moment because feeding them by adding media seems to encourage contamination? I use 1% P/S with the ATCC media. Do you ever completely remove the media? Is there another antibiotic you recommend? Many thanks
Dear Mª Carmen Ocaña and Denise Grana, I'm having problems with culture of this line at the moment because feeding them by adding media seems to encourage contamination? I use 1% P/S with the ATCC media. Do you ever completely remove the media? Is there another antibiotic you recommend? Many thanks
Hi, I am culturing the THP-1 cells yet I found that every time when I centrifuge them at no matter 200xg or 1000 rpm for 5 -10 min, I only have a pitiful tiny pellet. And there are still lots of cells that can still be observed in the supernate. Have you experienced this situation? Should I further increase the speed of the centrifuge or elongate the time? Will either of these cause cell lysis? Thanks!
Hi, I am culturing the THP-1 cells yet I found that every time when I centrifuge them at no matter 200xg or 1000 rpm for 5 -10 min, I only have a pitiful tiny pellet. And there are still lots of cells that can still be observed in the supernate. Have you experienced this situation? Should I further increase the speed of the centrifuge or elongate the time? Will either of these cause cell lysis? Thanks!
Hello I have a problem with THP-1 cell lines. I was working with them for more than 2 months without problem but recently every de frozen cell gets clump but grows well. Do you think what is the problem? Mycoplasma? if yes what is the reason and how I can find out and remove that? There are also some dot things, they are dead cells? Can we check that with Dapi staining? Thank you so much
Hello I have a problem with THP-1 cell lines. I was working with them for more than 2 months without problem but recently every de frozen cell gets clump but grows well. Do you think what is the problem? Mycoplasma? if yes what is the reason and how I can find out and remove that? There are also some dot things, they are dead cells? Can we check that with Dapi staining? Thank you so much
Dear Olivia, I did change media completely sometimes but I never did it because of contamination. I've never experienced it actually. I normally used RPMI 1640 and P/S 1% and it was perfectly suitable for me, thus I am not able to recommend any other antibiotic (just because I've never used another one)..! Adding media should preserve the substances released by the growing cells but should not allow or boost contamination (as far as I know).
Dear Olivia, I did change media completely sometimes but I never did it because of contamination. I've never experienced it actually. I normally used RPMI 1640 and P/S 1% and it was perfectly suitable for me, thus I am not able to recommend any other antibiotic (just because I've never used another one)..! Adding media should preserve the substances released by the growing cells but should not allow or boost contamination (as far as I know).
I need to start working with this cell line. I´ve found that cells should be seeded at 2-3x10^5 and split when reaching 8x10^5. Other suggest higher densities for seeding, splitting, and freezing. What densities would you recommend for seeding, splitting, and freezing? I will probably work on T25 flasks.
Thank you very much!
I need to start working with this cell line. I´ve found that cells should be seeded at 2-3x10^5 and split when reaching 8x10^5. Other suggest higher densities for seeding, splitting, and freezing. What densities would you recommend for seeding, splitting, and freezing? I will probably work on T25 flasks.
Thank you very much!
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I forgot to answer! Thank you a lot ! I will try doing it this way and hope it works well for me too =)
I forgot to answer! Thank you a lot ! I will try doing it this way and hope it works well for me too =)
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Dear Mª Carmen Ocaña and Denise Grana, I'm having problems with culture of this line at the moment because feeding them by adding media seems to encourage contamination? I use 1% P/S with the ATCC media. Do you ever completely remove the media? Is there another antibiotic you recommend? Many thanks
Dear Mª Carmen Ocaña and Denise Grana, I'm having problems with culture of this line at the moment because feeding them by adding media seems to encourage contamination? I use 1% P/S with the ATCC media. Do you ever completely remove the media? Is there another antibiotic you recommend? Many thanks
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Hi, I am culturing the THP-1 cells yet I found that every time when I centrifuge them at no matter 200xg or 1000 rpm for 5 -10 min, I only have a pitiful tiny pellet. And there are still lots of cells that can still be observed in the supernate. Have you experienced this situation? Should I further increase the speed of the centrifuge or elongate the time? Will either of these cause cell lysis? Thanks!
Hi, I am culturing the THP-1 cells yet I found that every time when I centrifuge them at no matter 200xg or 1000 rpm for 5 -10 min, I only have a pitiful tiny pellet. And there are still lots of cells that can still be observed in the supernate. Have you experienced this situation? Should I further increase the speed of the centrifuge or elongate the time? Will either of these cause cell lysis? Thanks!
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Hello
I have a problem with THP-1 cell lines. I was working with them for more than 2 months without problem but recently every de frozen cell gets clump but grows well. Do you think what is the problem? Mycoplasma? if yes what is the reason and how I can find out and remove that? There are also some dot things, they are dead cells? Can we check that with Dapi staining? Thank you so much
Hello
I have a problem with THP-1 cell lines. I was working with them for more than 2 months without problem but recently every de frozen cell gets clump but grows well. Do you think what is the problem? Mycoplasma? if yes what is the reason and how I can find out and remove that? There are also some dot things, they are dead cells? Can we check that with Dapi staining? Thank you so much
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Dear Olivia, I did change media completely sometimes but I never did it because of contamination. I've never experienced it actually. I normally used RPMI 1640 and P/S 1% and it was perfectly suitable for me, thus I am not able to recommend any other antibiotic (just because I've never used another one)..! Adding media should preserve the substances released by the growing cells but should not allow or boost contamination (as far as I know).
Dear Olivia, I did change media completely sometimes but I never did it because of contamination. I've never experienced it actually. I normally used RPMI 1640 and P/S 1% and it was perfectly suitable for me, thus I am not able to recommend any other antibiotic (just because I've never used another one)..! Adding media should preserve the substances released by the growing cells but should not allow or boost contamination (as far as I know).
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Nattan Stalin you may stimulate them with PMA. They will differentiate from monocytes to macrophages and adhere to the surface of the flask
Nattan Stalin you may stimulate them with PMA. They will differentiate from monocytes to macrophages and adhere to the surface of the flask
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Fan Zhang Hi, I always centrifuge 300g for 5 mn and it is working.
Fan Zhang Hi, I always centrifuge 300g for 5 mn and it is working.
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Gizem Eylem Baslar Thank you!!
Gizem Eylem Baslar Thank you!!
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Dear expertise,
How to prepare adherence cell line from THP-1 suspension cell line ?
Dear expertise,
How to prepare adherence cell line from THP-1 suspension cell line ?
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