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How to culture HK-2 cells


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#1 cell 293

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Posted 23 May 2013 - 07:17 PM

If i want to culture HK-2 cells, what medium i need to use.

How to prepare the medium?

#2 leelee

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Posted 24 May 2013 - 09:05 AM

Who did you get the cells from? What were they culturing them in, because this is what I would use.

Also, the ATCC is a good source of this type of information. The HK-2 listing can be found here:
http://www.atcc.org/...64650D34DF19048

I'm not sure what you mean by "how to prepare" the media?

#3 Tabaluga

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Posted 24 May 2013 - 01:49 PM

About the 'how to prepare': When you know the ingredients of your medium (like from the tab "Culture method" of the link above), you just mix them together and take note of possible extra instructions (like the one from that website, that the medium shouldn't be filtered).
Also, the described medium contains EGF, so I think the medium should not be warmed quickly in a waterbath but acclimatise at room temperature instead (At least our lab always handles it like that when a medium contains growth factors, please correct me if you think this is not standard/not necessary ?!)

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La chose simplement d'elle-même arriva,
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#4 bob1

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Posted 24 May 2013 - 05:36 PM

Also, the described medium contains EGF, so I think the medium should not be warmed quickly in a waterbath but acclimatise at room temperature instead (At least our lab always handles it like that when a medium contains growth factors, please correct me if you think this is not standard/not necessary ?!)

That's pretty standard, the amount of time warm should be kept to a minimum, and I think these go off quite quickly, so probably best to make them up in aliquots on the day of use if possible. Aliquots could be frozen with no loss of activity.

#5 leelee

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Posted 24 May 2013 - 07:59 PM

Still a little unclear on what part you don't understand sorry, but here is what I do with my media:

- purchase pre made media (e.g. DMEM) from a company
- purchase sterile additives (e.g. NCS, non essential amino acids, L-glu etc) at a stock concentration
- prepare and filter sterilise any further additives that I may need

- add the additives to my media to the desired concentration, store at 4C in the dark

- before use, remove bottle from fridge and allow to warm slightly at RT for 30 min or so- my cells don't mind cool media so I never bother with heating it in a water bath (other cells may be fussier). Also, water baths give me the heebie jeebies and are a contamination risk I am far happier to avoid :)


- I go through about 500ml a week to a fortnight so I don't bother with aliquoting, but if I used less, I would make up smaller aliquots as I go rather than the whole 500ml bottle at once.

Hope that helps :)

#6 cell 293

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Posted 26 May 2013 - 05:06 PM

Thanks for everyone's replications.

Sorry, i didn't describe this question clearly.

That is the culture medium what i use
- DMEM 500ml
- 3% L-glutamine 5.5ml
- Penicillin/streptomycin 5.5ml
- Q-FBS 50ml

Someone told me that is not suitable to HK-2 cells.

But the HK-2 cells looks regular by this medium.

Do i need change the medium what i use?

#7 bob1

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Posted 27 May 2013 - 01:59 AM

OK - follow Leelee's first post to start with - where did you get them and what have they been cultured in, in the past?

If you know this, then stick with that medium or wean the cells onto your preferred medium, but note that weaning should be carefully monitored to ensure that you aren't making any changes in the cell behaviour and responses to your experiments!.




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