Tagged Questions

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

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1answer
16 views

Cells “grown in LB broth to an OD600nm” [on hold]

Cells "grown in LB broth to an OD600nm" What does the OD stand for in this?
4
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2answers
37 views

Does trypsin strip flask coating?

Mammalian cell/tissue cultures sometimes require flasks coated with proteins. My uneducated guess is that these proteins mimic the ECM, perhaps the basal lamina, so finicky contact-dependent cells can ...
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0answers
16 views

Best method to culture lung fibroblasts?

I am just looking for the best method to culture human lung fibroblast from a needle biopsy?
8
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1answer
227 views

What's the difference between growing cells in culture and cloning them?

The wikipedia page on Hela cells refers to George Gey being able "to isolate one specific cell, multiply it, and start a cell line." Later it says, "In 1955 HeLa cells were the first human cells ...
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0answers
18 views

Creating Sperm via cell culture adaptation

I am wondering if it is theoretically possible to create "artificial sperm" via cell culture adaptation. That is at very high level, can you replace the genes of one sperm cell with those of another ...
3
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1answer
49 views

Looking for protocol to produce a drosophila primary cell line

I'm looking for a general (and easy, this is going to be for an undergrad class) protocol to isolate and culture primary cells from fruit flies.
2
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1answer
52 views

How do I interpret this graph regarding introduced genes and virus-infected cells?

This graph appeared in a practice test for the MCAT. I am trying to interpret it, but it confuses me a bit. On the x-axis we have some introduced genes, and on the y-axis we have % of cells ...
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0answers
25 views

Experimental studies of bacterial adaptation to changes in nutrients of culture medium

Are there experimental studies that measure how a population of bacteria in continuous culture evolve when submitted to changes in the concentrations of metabolites in the culture media? For example, ...
1
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1answer
164 views

RNeasy Mini Kit low 260/230 ratio — can I purify this RNA for further use?

I used Qiagen's RNeasy Mini Kit to isolate RNA from 5*10^5 C28/I2 (immortalized human chondrocytes). However, my yield is low (~25 ng/ul), but my 260/280 ratio is great (~2.3), and my 260/230 ratio is ...
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0answers
32 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
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1answer
81 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
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1answer
57 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 ...
8
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2answers
409 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 ...
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0answers
31 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 ...
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0answers
26 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
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1answer
76 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. ...
5
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1answer
153 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?
2
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0answers
104 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
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2answers
112 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 ...
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1answer
44 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
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1answer
49 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
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1answer
150 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 ...
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2answers
123 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 ...
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2answers
596 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 ...
7
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1answer
234 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, ...
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0answers
40 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 ...
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0answers
58 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
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1answer
957 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 ...
4
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1answer
46 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
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1answer
111 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
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2answers
679 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 ...
3
votes
1answer
165 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 ...
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0answers
169 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
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1answer
232 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 ...
2
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1answer
431 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 ...
4
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2answers
2k 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
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0answers
124 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 ...
5
votes
1answer
981 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
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1answer
79 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:
4
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1answer
168 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 ...
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0answers
330 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 ...
9
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2answers
9k 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
3k 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
votes
1answer
55 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 ...
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5answers
630 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 ...
5
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0answers
115 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
80 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
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0answers
34 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
157 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.