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

learn more… | top users | synonyms

0
votes
0answers
21 views

Cells and their characteristic properties

Could a culture of cells of a certain type be 'forced' to attain certain properties in successive generations that would have been harmful to the cell culture as they are now?
2
votes
1answer
45 views

Are cell lines potentially dangerous?

More specifically, if a human subject was exposed to, say, a human cancerous cell line (via intravenous injection or through an open wound, for example), is it possible that they would develop any ...
3
votes
1answer
30 views

Hela live cell confocal laser scanning - reccommendations for good fluorophore that will show good movement

I've been doing a lot of live cell imaging lately mostly using hela cells expressing some EYFP based chimeric proteins. I'm building a video library for an art student here at the university who is ...
2
votes
1answer
26 views

Solubility of Forskolin in ethanol

I am interested in using forskolin in cell culture medium. Does anyone know how to make solution of 10 microM forskolin in 5% ethanol or less. I would like to avoid using DMSO as a solvent. Thank you. ...
6
votes
2answers
268 views

Gloves for Cell Culture?

I always wear gloves when I'm doing cell culture. Moreover, I always spray my gloves with ethanol to disinfect, so that I don't contaminate everything. However, I recently heard the argument that ...
0
votes
0answers
21 views

How to start Diagnostics of Enterobacteriaceae?

I am thinking what is the Diagnostics for Enterobacteriacea and particularly Shigella, Yersinia, Vibrio, Campylobacteriacea, Helicobacter pylori, salmonella and Proteus. I got today advice that start ...
0
votes
0answers
19 views

Macrophage death by toxin. Little balls inside cell? What type of death?

After I treated J774 cells ( a mouse monocyte/macrophage cell line ) with Clostridium difficile Toxin B (1 ng/ml for 44 hours), I noticed several small circles inside of a seemingly dead cell. I'm ...
2
votes
0answers
45 views

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?
8
votes
2answers
6k 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?
2
votes
0answers
89 views

Are HeLa cells edible?

I'm curious if HeLa cells are intrinsically poisonous or dangerous to ingest. My understanding is that some of the contamination in HeLa cells such as HPVs are not readily expressed. I have no plans ...
4
votes
2answers
96 views

What is the smallest oligocelluar organism?

What is the smallest oligocelluar organism? How many cells does it have? EDIT The question is motivated by this comment@Philosophy.SE EDIT as recommended in comments I'm looking for an example of ...
2
votes
1answer
196 views

Where to put the gene after eukaryotic promoter for best expression levels?

As far as I know there is an optimum distance between a promoter and the gene for the best expression levels. What is that distance for common promoters like CMV, SV40? If you have a first hand ...
0
votes
1answer
28 views

Looking for raw cell videos from a light microscope

First off, I'm not looking for online lectures. I am trying to find creative commons licensed videos of live cells moving around without annotation or anything extra. I would prefer a few minutes of ...
1
vote
1answer
45 views

The reason why researchers usually use cell lines from “blast cells”?

What's the reason why researchers usually use cell lines from "blast cells" (so, immature, like lymphoblastoid cells) for measuring gene expression data? Is that they are growing up, which would make ...
5
votes
1answer
112 views

Measuring algae growth or death

My daughter (10th grade) is doing a science fair project on the toxicity of triclosan for the algae Selenastrum capricornutum. She is wondering the best way to measure the effect, given the limited ...
8
votes
2answers
723 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, ...
4
votes
2answers
475 views

How does formaldehyde/PBS or methanol fixation of cells affect lysosomal pH?

The question is fairly simple - does formaldehyde or methanol fixation in preparation for immunocytochemistry/immunofluorescent staining affect the pH of the lysosomes? Some background: I'm trying to ...
0
votes
2answers
86 views

How is it that the WI-38 cell line isolated by Hayflick in 1962 is still very much around and not affected by the 'Hayflick Limit'?

I have searched the net and I have not been able to come up with an clear answer. Edit: Here is the para quoted from Nature http://www.nature.com/news/medical-research-cell-division-1.13273 "So began ...
3
votes
0answers
153 views

How to reduce/eliminate cell clumping in suspension CHO cells, without using an anti clumping agent?

what is the best way to eliminate clumping suspension cells used for transfection. Anti clumping agents interfere transfection and hence can't be used. Though maintaining the cell density low helps, ...
5
votes
0answers
91 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 ...
3
votes
1answer
136 views

What is the best way to clean plastic flasks that have been used for cell cultures - is virkon a good idea?

When you use cultures e.g. insect cells, which are infected with virus one way to clean the (plastics) shake flasks is with virkon. Which is the most effective way to clean your flasks in order to ...
0
votes
0answers
35 views

Generic Abbreviation for Immunopanning?

This is actually something I've been wondering for a while. I've seen all kinds of short hand used outside of papers (IPn most commonly), but in publications I've not seen an abbreviation for ...
0
votes
0answers
33 views

Freezing collagen aliquots

I'd like to regularly coat some dishes with collagen (Rat Tail, Sigma C7661) for use with PC12 cells. As I only need a small amount each time, I would like to know about any experience in freezing ...
2
votes
1answer
402 views

fibroblast cells and fibers

I am interested in fibroblast cells in human arteries. Here are the things that I am not clear at the moment and I could not find any answer from the literature: What are the dimensions of these ...
3
votes
0answers
32 views

How different are tissue-specific fibroblasts from each other?

I am planning to utilize a new system in our lab, in which I will co-culture cancer cells from different tissues with fibroblasts. I have the option to receive skin-derived primary fibroblasts. I ...
3
votes
1answer
83 views

How do different tissue culture matrices affect background in fluorescent microscopy?

In response to my previous question, I've been reading up a little bit on poly-D-lysine, Collagen I, Collagen IV, laminin, and other tissue culture coatings that promote cell adhesion. I've always ...
4
votes
1answer
392 views

How to convince suspension cells to adhere more tightly?

I'm developing a cell-based assay in 96-well plates that requires adherent cells, as they need to be washed at least twice during the protocol. I'm using in-house strains of HT1080 cells (some ...
1
vote
0answers
104 views

What mammalian cell lines are easiest to grow and transfect?

I'm working with HEK293 cells, which are very easy to transfect. Does anyone have other suggestions for cell lines with very high transfection efficiency? Ideally the chromosomes would also be diploid ...
2
votes
1answer
168 views

Copper for cell incubator to prevent contamination

For some reason the lab seems to have a problem with contamination every so often. It's virtually impossible to prevent bacteria, viruses, fungi, etc. from getting into the incubator every time you ...
3
votes
2answers
1k views

How can I resuspend a cell pellet without harming the bacteria?

When using a preculture with Ampicillin in my protein expression, I have to get rid of the preculture medium to avoid carrying over too much beta-lactamases that will destroy the ampicillin in the ...
2
votes
0answers
88 views

Resuspending Cells from a filter plate

My experiment involves pulling down cells on to a filter plate for a variety of assays. However, I want to ensure that my cells are alive so I am trying to resuspend them so that I can do an accurate ...
4
votes
1answer
140 views

Is cell senescence in culture comparable to that in vivo?

A cell is 'senescent' when is has permanently left the cell-cycle. This can be caused by stresses, or by reaching the 'Hayflick limit' (the cell has reached its replicative lifespan, as defined by its ...
5
votes
1answer
759 views

High Glucose vs Low Glucose DMEM for Cell Culture

I've noticed that in mammalian cell culture, there are often two types of DMEM available. High Glucose and Low Glucose. Does it matter which type I use for culturing of cells (e.g. Hela or HEK293)? ...
2
votes
1answer
69 views

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:
1
vote
0answers
283 views

Appropriate methods for triteration?

What is the appropriate method to pipetting up and down mammalians cells to sufficiently break up clumps. This should also keep in mind the issues from Pipetting damage on cells. Visually one can see ...
2
votes
1answer
49 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 ...
3
votes
1answer
2k 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.
4
votes
2answers
410 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 ...
7
votes
1answer
67 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 ...
5
votes
0answers
43 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 ...
3
votes
0answers
33 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?
5
votes
3answers
850 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.
7
votes
1answer
137 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 ...
6
votes
1answer
233 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 ...
24
votes
3answers
3k 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 ...