Biology Stack Exchange
Biology Stack Exchange

Questions tagged [cell-culture]

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

Cell culture is the process of taking living cells and growing them in vitro: under controlled laboratory conditions, isolated from their original organism and natural environment. Cells may be cultured directly from tissues extracted from the parent organism (known as "primary" culture), or from immortalised cell lines such as HeLa, of which a great variety are now commercially available.

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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 stock is troublesome). Some think using it…
Nick T
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Pipetting damage to cells

I'm curious how much damage is potentially inflicted by shear stress by pipetting. I know that syringes used for stem cell injection can cause a lot of damage. However, to what extent does this happen with P20 and P200 pipette tips? Understandably…
bobthejoe
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Microinjection intestinal organoids

I am attempting to microinject small intestinal organoids, using the Eppendorf FemtoJet 4i microinjector and Femtotip capillaries, and have come across several issues. This is a new set up in our lab, and I am first to attempt it so I apologise for…
Lemity
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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, when I did the same with INS1-E and MIN6 (both…
kali281
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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 express this specific receptor? Are there any…
kali281
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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.
bobthejoe
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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 that these cells should just be kept at room…
bobthejoe
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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 transfected with NR1 and NR2x subunit cDNAs and also GFP,…
walkytalky
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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.
Rob O'Callahan
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What's the recommended maximum passage number for mammalian cells (NCI-H441)?

Is there a generally accepted maximum number of times you should split and reseed mammalian culture cells before restarting cultures from new stock?
Dan Riggins
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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?
bobthejoe
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Are cryovials really better than regular tubes for cryopreservation?

What is the rationale for using cryovials instead of regular, sterile centrifuge tubes for the storage of mammalian cells? With a good aseptic technique and without touching the cap directly, there should not be a significant risk of contamination…
Mowgli
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Tubing connector identification request

this may not be on topic but I'm hoping someone here can tell me the name/manufacturer of the connectors used with the thin teflon tubing in this picture:
so12311
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Testing CO₂ for cell culture

I'm am refurbishing some old CO2 incubators. I would like to test the precision of the CO2 sensor. There is a sample port on this incubator. What sorts of methods are commonly used to measure CO2 in these incubators? Strangely, the manufacturer of…
rhill45
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What is the standard protocol for creating permanent stocks from freeze-dried bacteria?

I've received freeze-dried bacteria that I intend to use for a study I'm doing on slime mold; however, I've never dealt with freeze-dried, only frozen, bacterial cells. After reconstituting these cells in 6mL media, is it alright to split this in to…
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