12 replies to this topic

[epigenetics]

Hello,
We decided to go till 90% conflency level of human skin fibroblast cells. and  theres a problem, the pic one of my lab members showed me for 90% conflency, other one argued that it is 60-70%. My cells are getting ready for harvesting,  Experts, would you be able to show me any pic of cells at 90% confluency.
so that i can do it properly,
Thanks,

[Stephan]

hahaha, very nice.  I completely agree with you - everyone has a different idea of what looks like 60% or 70% etc.  I guess it depends largely if you are a glass half full or glass half empty kind of person.  Sorry I don't have any pics but interested to see any further posts with pics - may start an interesting discussion.

[little mouse]

I don't have any pic of subconfluent cells neither. post a picture of your cells and we will tell you what is the confluence.
(I guess you will have various responses from 70 to 90 !)

[epigenetics]

Here is the pic i have been shown for 90% confluency level of human skin fibroblasts

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[Alfred Nobel]

epigenetics, on Aug 6 2009, 01:17 PM, said:

Here is the pic i have been shown for 90% confluency level of human skin fibroblasts

those cells appear closer to 50 than 90%. When fibroblasts approach 90% you will see them in contact with one another and swirls usualy start to form. The plate will be almost entirely covered with only a few empty areas.

[epigenetics]

Alfred Nobel, on Aug 6 2009, 01:52 PM, said:

epigenetics, on Aug 6 2009, 01:17 PM, said:

Here is the pic i have been shown for 90% confluency level of human skin fibroblasts

those cells appear closer to 50 than 90%. When fibroblasts approach 90% you will see them in contact with one another and swirls usualy start to form. The plate will be almost entirely covered with only a few empty areas.

The pic is not coming clean, though my pic what i scanned is very clean. However, If you consider the cytoplasmic area inside, they might not be 50%, but more than that. Whats the percentage? i am not sure.

[bob1]

The problem with confluency is that it is different for different cell lines. I occasionally work with brain cell derived cells that have very long processes and are contact inhibited, so while only 30-40% of the apparent flask area is covered, the cells stop growing having reached 100% confluency by the processes touching.

In terms of area, your cells will be about 50-70%, though the picture is a little unclear and without knowing the cell line it is hard to judge how confluent they actually are.

[leelee]

I agree with those above that the picture you posted looks like between 50-70% confluent.

[miBunny]

Add one more vote for the 50-70% confluent!

[genehunter]

I would say its more like 60% to me.

[epigenetics]

bob1, on Aug 6 2009, 05:24 PM, said:

The problem with confluency is that it is different for different cell lines. I occasionally work with brain cell derived cells that have very long processes and are contact inhibited, so while only 30-40% of the apparent flask area is covered, the cells stop growing having reached 100% confluency by the processes touching.

In terms of area, your cells will be about 50-70%, though the picture is a little unclear and without knowing the cell line it is hard to judge how confluent they actually are.

Thanks to all of you for sharing your idea. this is one patient's (Human) skin fibroblast what we have grown at lab.

[Stephan]

Hard to tell the confluency, I wouldn't say it's 90% though.

I think you need to determine it experimentally - see how dense they will grow and once they have reached that point you know what 100% looks like.  Then just go a little less I guess.

[rhombus]

epigenetics, on Aug 5 2009, 09:01 PM, said:

Hello,
We decided to go till 90% conflency level of human skin fibroblast cells. and  theres a problem, the pic one of my lab members showed me for 90% conflency, other one argued that it is 60-70%. My cells are getting ready for harvesting,  Experts, would you be able to show me any pic of cells at 90% confluency.
so that i can do it properly,
Thanks,

The only true way of estimating the confluency is to use a Graticule and count the number of cells in a field. If you grow cells to what you think is 100% and count with a graticule, then subsequent measures of confluency are easy.

Of course every cell/line is different (size and shape), so you would have to do this process with every cell you use.

Kindest regards

Rhombus