Cell culture is the process of growing and maintaining living cells in vitro.

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3answers
12k views

Hoechst or DAPI for nuclear staining?

When is it best to use Hoechst vs. DAPI for nuclear staining? They seem to be very similar on paper. Are there situations where one is clearly preferable?
3
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1answer
4k views

Why is beta-mercaptoethanol often added to cell culture media

Many protocols suggest that beta-mercaptoethanol is necessary for growing cells. It is a reducing agent but what does it mechanistically do for your cells. When would one not add it.
2
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1answer
60 views

What is an efficient way to spike 10-50 cells to a culture

I'm trying to add on the order of 10-50 mammalian cells to a mixture of other cells (order of 1e6 cells). What is the best way to do this? (edit) I guess this wasn't clear. I would really like to ...
17
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5answers
886 views

Pipetting damage on cells

I'm curious how much damage is potentially inflicted by shear stress by pipetting. I know that with syringes for stem cell injection cause a lot of damage. However, to what extend does this happen ...
6
votes
1answer
156 views

What temperature should mammalian B-Cells be stored at outside of the incubator?

I'm working with murine B-cells. The general protocol is to keep cells on ice to keep them from dying but I've noticed that it makes these cells aggregate and precipitate out. I've heard suggestions ...
7
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1answer
93 views

Providing small molecules to cells on a filter plate

Lets imagine that I have mammalian cells that I've immobilized on a filter. Now I want to keep providing small molecules to these immobilized cells without resolubilizing the cells. The caveat is ...
4
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1answer
45 views

Can a cell culture insert plate be used with a vacuum manifold?

There exists 96 well plate setups for use with vacuum manifolds. Is it possible to use 6/12/24 plate inserts with the same setup?
7
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1answer
168 views

Which human cell lines do not express the GLP-1 receptor?

I need a human cell line that does not express the GLP-1 (glucagon like peptide-1) receptor. I'm working with HeLa cells, do those express the GLP-1 receptor? Which other cell lines exist that don't ...
5
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3answers
1k views

How do you break up cell clumps when passaging?

I am working with HepG2 cells, and they really like to form clumps. Pipetting up and down in TrypLE does not seem to be very effective in breaking them up.
8
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1answer
71 views

What is the effect of exendin on beta-cells

Do you know if exendin, an analog of GLP-1 (glucagon like peptide-1), can be toxic for beta-cells? For example what is the effect on INS1 or Min6 cells at a certain concentration or after 90 mins of ...
8
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2answers
1k views

How much time do INS1-E and MIN6 cells need after splitting?

I am currently doing an experiment on cells to test the internalization of a protein. Normally, I seeded my cells the day before the incubation. This worked well for Hela, CHL or PANC1 cells. However, ...
7
votes
1answer
341 views

How can I keep HEK cells alive while expressing NMDA receptors?

I am trying to express functional NMDA receptors in HEK293 line cells for single channel recording experiments. The HEK cells are maintained in the standard way (Thomas & Smart 2005) and ...
26
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3answers
5k views

How long does antibiotic-dosed LB maintain good selection?

Various people in our lab will prepare a liter or so of LB, add kanamycin to 25-37 mg/L for selection, and store it at 4 °C for minipreps or other small cultures (where dosing straight LB with a 1000X ...