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SH-SY5Y and Poly-L-Lysine


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9 replies to this topic

#1 NDUFB11

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Posted 17 May 2013 - 05:21 PM

Hello everybody,
I am having problem with neuroblastoma cell line, in particular when I seed the cells on the petri dish, most of them remain in suspension.
Looking on line I see that to overcome this problem some people use poly-l-lysine but I have no idea how to use it,
furthermore, sigma has many type of poly-lysine, could please someone tell me how to use it ? and which kind of poly-lysine is good in term of molecular weight ?

Regards

Patrizio

#2 bob1

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Posted 18 May 2013 - 12:44 AM

It doesn't matter too much which one you use, so long as it says cell culture tested or cell culture grade. Protocol here, which will probably recommend a product, seeing as it is a Sigma protocol.

#3 h9494036

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Posted 18 May 2013 - 06:51 AM

I have used addexbio coat, and it works wonderfully for neuronal cells, they are cell culture ready, just dilute it to 1x and should do. here is the link: http://addexbio.com/...detail?pid=4666

Edited by h9494036, 18 May 2013 - 06:51 AM.


#4 NDUFB11

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Posted 18 May 2013 - 08:14 AM

Thank you guys
I will try to use that soon

#5 bob1

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Posted 18 May 2013 - 05:43 PM

I'll add that SH-SY5Y normally consist of a mixed population of floating and attached cells - you need to ensure that both populations are maintained when culturing.

#6 BRD

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Posted 20 May 2013 - 08:25 AM

If attachment is the issue, you may want to try these vessels designed for improved attachement of cell lines:

http://www.corning.c...nhanced_tc.aspx

It's easier and cheaper than dealing with Poly-l-lysine

#7 NDUFB11

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Posted 25 May 2013 - 11:39 AM

I tried using the sigma product P1399-25MG, I followed the steps on their protocol but for some reason it didn't worked as I aspected,
attached is the picture,

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Edited by NDUFB11, 25 May 2013 - 11:42 AM.


#8 NDUFB11

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Posted 25 May 2013 - 11:41 AM

as you can see in this image, after 24 hours most of the cells are attached, but they appear spherical and weird, maybe there is some step that needs to be modified or anyway I don't know ...if you guys have any suggestions could be useful for me and for somebody else in the future

thank you

#9 h9494036

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Posted 14 September 2013 - 05:49 AM

AS I recall using SH-SY5Y before, I did not need to use poly-lysine as long as you use a cell-culture treated flasks. Your cells might have some issues.

 

#10 scolix

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Posted 14 September 2013 - 11:42 AM

Plate higher number of cells, and use cell culture treated plates. these help with SHSY5Y cells. 






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