Is anyone here culturing H9C2 cells? My H9C2 cells growth rate is very slow and I am not sure what is the problem with this. I am using DMEM with 10% FBS and 1% sodium pyruvate and Passage 24 of H9C2 cells. Some info from this website say that we have to do a clone selection by using a 96 well plates. Can anyone share how to do this?
Finally I am done successfully of cone selection of H9C2 by just doing trial and errors. Anyone want to know, just raise your queries.
Finally I am done successfully of cone selection of H9C2 by just doing trial and errors. Anyone want to know, just raise your queries.
No worries as you are getting a new stock from ATCC right. Pretty easy. The most important is do not wait until cells are confluent about 80-90 or 100%. Harvest when it reaches to 70 to 75%. If you keep till 100%, you will see some changes of morphology.
You need very low amount of trypsinEdta. It works with very low concentration too eg. 0.05% (original bot comes with 0.5% of trypsin). Dilute with PBS to get 0.05%. Make sure you get single cell suspension to pass them for sub-culture. Hope this works well.
Use DMEM and 10%FBS.
Hi Santosh, I did not try transfections. Yes I did making stable for it. Don't wait till cells became 80 to 90 % confluent. Better sub-culture earlier. This is key for this H9C2. Since your one is fresh from ATCC, you no need to worry much. All will be great.
Later after you might notice that you need to do clone selection when you see cell growth rate become noticeably slow (not so soon and not worry much). Important thing is you just observe cells characters daily and make sure you are familiar with them so that any changes you know straight away.
NaP' is not necessary to add if your media is fresh. You may consider to add it if your media is old. DMEM, 10% FBS and antibiotics works well. Hope this helps.
Ming Chuan Yu
Can you tell me why extracellular matrix is coming in H9c2 cells so fast ? Why my MTT result is so fluctuated in H9C2 cells line ?
Hi Islam...Can you elaborate more accurately for your Questions. Indicate your extracellular matrix in your picture and you have to tell me how you do your MTT assays. Without saying how you did your assay, no one can answer for it except yourself.
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