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

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Is there any alternative to cryopreserve cell lines without -80 freezer?

My university's -80 freezer just broke down and the repair will take 4 months according to the vendor. Is there any other way to preserve cells? Maintaining lot of flasks until the freezer is repaired ...
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Cationic lipid mediated transfection optimization for 150mm dishes

I need to optimize a transfection protocol to transiently express a plasmid encoding a chimera of eyfp attached to the c term of a Golgi apparatus signaling molecule) in hela cells and hepg2 cells ...
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What type of flask should I use to culture NTERA2 embryonic cancer stem cells?

I'm just starting my MSc research and I am in the process of making a list of equipments/consumables to order. Is there a specific flask in which I can culture NTERA2 (NTERA2/D1) cell line?
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alternative ways to detach cells other than trypsin [duplicate]

So I have an A549 (rather "sticky" cells, they usually take about 7 minutes to trypsinise) cells and I cannot use trypsin or tripLE as this will skew my ELISA results. I have tried scraping the cells ...
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166 views

What does confluency mean?

Since as long as I have been doing cell culture, the word confluency is used to describe the % growth of cells or area covered by them. However, no dictionary that I have found uses this word. I was ...
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What microscope/magnification would I need to observe P. Acnes bacteria?

I am currently attempting to grow a culture of P. Acnes bacteria. Right now, my only hope in identifying colonies of the bacteria in the culture is to use a black light to find colonies that glow ...
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43 views

Why are cell lines frozen in vapor phase?

For most of the cell lines I've come across, ATCC recommends storing them in the vapor phase of liquid nitrogen. I'm taking that to mean any place above the level of liquid in the nitrogen storage ...
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19 views

Cells scatter light in solution. Do they refract?

For part of an exam question, "why do cells grown in liquid cause turbidity?" I answered that cells refract light. The correct answer was that cells scatter light. Isn't the light scattering ...
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37 views

Estimating RPM to RCF in Methods from Older Papers

I'm attempting to replicate a cell biology method from a 1958 Laboratory Investigation paper. The protocol is for the isolation of an extracellular matrix protein, and a key step is a centrifugation ...
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123 views

Studying effects of alcohol on cells

I am wondering about the logistics of a simple experiment with say 3 types of alcohols, and various concentrations of each, with a control(s) I would like to research the effects of alcohol on ...
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How does air-liquid interface (ALI) culture work?

For studying diseases, such as chronic obstructive pulmonary disease (COPD), cells are often grown in air liquid interface. I understand that the common way of establishing this these days is to grow ...
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Henrietta Lacks and HeLa cells

Why did Henrietta Lacks die when her cells didn't die? As I understand it only the cells from her cervix were immortal so perhaps the cervical cancer took her life and the cells never died.
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Pink E.coli cell pellet. Reasons? [closed]

I've harvested IPTG induced E.coli BL21DE3 cells' suspension culture by spinning at 5000 rpm/15 min/4 degrees C. The pellet after spin looked pale pink colored. What could be the reasons?
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What does it mean for the signals for transcription and translation to be “conserved”?

I was reading this article: "Overview of vector design for mammalian gene expression." for an explanation of why mammalian cell lines are used for expressing cloned genes, and one of the reasons ...
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Effect of flask position in shaking incubator affects growth kinetics

During some cultivation experiments for my bachelor thesis I noticed that in liquid culture my bacteria (E. coli) grew differently, according to the position in the shaking incubator. Because the ...
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42 views

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

Cells "grown in LB broth to an OD600nm" What does the OD stand for in this?
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181 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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280 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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21 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 ...
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65 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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110 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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562 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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162 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 ...
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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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521 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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120 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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297 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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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 ...
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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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77 views

Plant tissue culture [closed]

Tissue culture means that cells are propagated and multiplied under controlled conditions. Suppose the following situations: a) A seed being is embedded in an Agar medium and germinated under ...
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54 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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54 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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183 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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164 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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736 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 ...
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451 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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2k 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 ...
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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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139 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 ...
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882 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
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187 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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281 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
679 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 ...
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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 ...
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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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84 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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198 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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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?
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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.