Citing Robert Insall <[log in to unmask]>:
> Harry Macwilliams used to make conditioned medium by growing cells
> at a great density for a relatively short time. He then spun out
> the feeder cells and used the conditioned medium to initiate growth
> arrest in fresh cells. This got over the problems Rich describes,
> but I don’t know how well it works in practice.
>
> Nocodazole makes cells very, very sick, so the value of that
> syncronization depends on how physiologically normal you need them
> to be, and how soon after release.
>
> I toyed with the idea of making a Shokat mutant in wee1 or a
> nondegradeable cyclin B, but it hasn’t been something I needed
> enough….
>
> R
>
>
> On 15 Sep 2016, at 15:09, Richard Gomer
> <[log in to unmask]<mailto:[log in to unmask]>> wrote:
>
> Hi-
> One way is to grow the cells to stationary phase, let them sit there
> for 1 day, and then dilute them, the polyphosphate in the
> conditioned medium causes a G2 arrest. This method has variable
> results, as being at stationary for too long does odd things to the
> cells, so I've found that this works 1 in 3 times or so. Tto verify
> the synchronization, you count the cells in the released population
> once an hour. If there is a steady increase, the synchronization of
> course didn't work; if the density is flat for 4-6 hours, and then
> suddenly jumps by a factor of 1.6-2, it worked..... Another way is
> nocodazole (microtubule disruption), this locks them in M phase, and
> then you wash the cells to release them into the cell cycle.
> best,
>
> Richard Gomer
> Thomas Powell '62 Professor of Biology
> Texas A&M University
> ILSB MS 3474
> 301 Old Main Drive
> College Station, TX 77843-3474
> 979 458 5745
>
> ________________________________________
> From: DICTY
> [[log in to unmask]<mailto:[log in to unmask]>] on behalf of Carl Franck
> [[log in to unmask]<mailto:[log in to unmask]>]
> Sent: Thursday, September 15, 2016 8:33 AM
> To:
> [log in to unmask]<mailto:[log in to unmask]>
> Subject: [DICTY] Please advise as to how to at least partially
> select Dicty in suspension in some phase of the cell cycle
>
> Dear Dicty Community,
> We were hoping someone might have a suggestion for how we can to
> some degree get a suspension culture readily into cell cycle
> synchronization. Years ago, an anon referee warned that turning room
> lights on and off matters. Much obliged for any advice! Sincerely,
> Kimberly Chen and Carl Franck, Ithaca, New York USA
>
>
> --
> Carl Franck
> Associate Professor, Physics
> Clark Hall E18, 525
> Cornell University, Ithaca, NY, USA
> website:
> https://urldefense.proofpoint.com/v2/url?u=http-3A__franckgroup.lassp.cornell.edu_&d=CwIFAg&c=yHlS04HhBraes5BQ9ueu5zKhE7rtNXt_d012z2PA6ws&r=G0i-xkKvWepiOT01FF_Nx9XkaEEFt5Dttsc3yIePxBPU44aHfHsMfVSlUygwIJiN&m=kPtLlGKrhd_OxZnv5QCGCwOsLS8BoOG4Ruq1RALTMSQ&s=giQ_iNYCOTKqxDKLz_BlnTVrZIYwM7SDk0FMah76k9Q&e=
> <https://urldefense.proofpoint.com/v2/url?u=http-3A__franckgroup.lassp.cornell.edu_&d=CwMFaQ&c=yHlS04HhBraes5BQ9ueu5zKhE7rtNXt_d012z2PA6ws&r=G0i-xkKvWepiOT01FF_Nx9XkaEEFt5Dttsc3yIePxBPU44aHfHsMfVSlUygwIJiN&m=5bUHbDBF_ljdOt_t8lYFAShaxVqwWaWvxyPThw5Dv_0&s=-1FZdKTM-o-Ac3ooN67zHD6FX9_xUqCtGWdtSA2nqu0&e=>
> lab phones: 607-255-5215<tel:607-255-5215> and 255-3562 (long ring
> for messages)
> home phone: 257-6969 (until 10PM)
> cell phone: 607-229-7190<tel:607-229-7190> (sometimes unreachable
> in my lab, if so, please use above lab phone number)
>
> --------------------------------------
> Professor Robert Insall
> Cancer Research UK Beatson Institute
> Switchback Road, Bearsden
> Glasgow G61 1BD, UK
>
> Tel. (0/44) 141 330 4005
> Web
> https://urldefense.proofpoint.com/v2/url?u=http-3A__www.beatson.gla.ac.uk_robert-5Finsall&d=CwIGaQ&c=yHlS04HhBraes5BQ9ueu5zKhE7rtNXt_d012z2PA6ws&r=G0i-xkKvWepiOT01FF_Nx9XkaEEFt5Dttsc3yIePxBPU44aHfHsMfVSlUygwIJiN&m=aWoCcrMo0-icGava06Mp7t_Kw8EagxXpEA5Kjc-u8ZQ&s=BvZ7HlDaoCr7OCD7LSgT8KSFkKaD6jZIwi4LZYAlea0&e=
> Orcid 0000-0003-4898-040X
> --------------------------------------
>
>
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Prof. Igor Weber, PhD
Ruder Boskovic Institute
Dept. Molecular Biology
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Fax: ++385-1-4561177
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