Culturing HeLa Cells - (Feb/22/2010 )
Hi Guys,
I am very new to culture of mammalian cells and I have to design a protocol as a part of my Lab work.Can any one explain me how do we start a culture of HeLa cells from frozen cells.Which would be the best culture medium for Hela Cells ? Are these cells adherent or in suspension while in culture ?
Secondly , I will be transfecting these cells using three different techniques in 24-well plate. So, I would like to know
about transferring these cells to the 24-well and how to maintain them during transfection, how often to change the medium. Do we bring the cells in suspension before transfection ?
I have read several protocols but I have not very well understood. I hope you can make out from my questions that I have no experience of culturing mammalian cells.
Kindly help me.
Thank you.
Your best bet is to find someone who can show you the basics of cell culture rather than reading through protocols.
Having said that, you might find a book like R.I Freshney's Culture of Animal Cells: a laboratory manual to be useful. It has all the methods you need to get you started.
For your questions... Have a look at the ATCC (American type culture collection) entry on HeLa, which gives basic culture conditions. Be aware that what you culture them in should be the same medium as the person who froze them down used, you can wean them onto other media at a later time if you like. HeLa are very tough and will survive a wide range of conditions apart from those on the ATCC website. HeLa are adherent.
For the seeding into plates, first learn how to count cells on a haemocytometer.... From your counts you can work out how many cells you have per volume, which can be used to work out the dilutions required. If you are used to doing dilutions in the lab, this will be no problem, use the same formulas. Roche Lab Faqs has useful data like cell numbers per well/plate/flask that you will find useful. There are heaps of threads on here that explain in detail how to count and dilute cells.
bob1 on Feb 22 2010, 04:30 PM said:
Having said that, you might find a book like R.I Freshney's Culture of Animal Cells: a laboratory manual to be useful. It has all the methods you need to get you started.
For your questions... Have a look at the ATCC (American type culture collection) entry on HeLa, which gives basic culture conditions. Be aware that what you culture them in should be the same medium as the person who froze them down used, you can wean them onto other media at a later time if you like. HeLa are very tough and will survive a wide range of conditions apart from those on the ATCC website. HeLa are adherent.
For the seeding into plates, first learn how to count cells on a haemocytometer.... From your counts you can work out how many cells you have per volume, which can be used to work out the dilutions required. If you are used to doing dilutions in the lab, this will be no problem, use the same formulas. Roche Lab Faqs has useful data like cell numbers per well/plate/flask that you will find useful. There are heaps of threads on here that explain in detail how to count and dilute cells.
Thank you, the links you have suggested have helped. Actually I have no one who can show me how to the basics of cell culture, but I am trying to equip myself the most before starting the work in lab. It is a self-to do exercise.