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

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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 ...
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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 ...
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532 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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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 ...
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96 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.
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209 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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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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850 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?
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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 ...
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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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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 ...
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1answer
215 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. ...
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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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178 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
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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 ...
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1answer
66 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 ...
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1answer
59 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 ...
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377 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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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, ...
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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 ...
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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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239 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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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 ...
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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 ...
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1answer
372 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 ...
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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 ...
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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
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1answer
233 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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1answer
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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)? ...
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97 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:
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1answer
79 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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1answer
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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.
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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.
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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 ...
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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 ...