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

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How to validate the use of an anti-virus monoclonal antibody in IHC by spiking a fresh organ with infected cultured cells?

I have assessed the specificity of a particular monoclonal antibody against a virus by immunofluorescence. I'd like to further validate it by testing it by IHC. Therefore I'd like to infect cells in ...
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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 ...
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Cell line immortalisation biosafety

Cell lines are often immortalised by artificial expression of proteins which specifically cause the knockout of important cancer suppression genes or the activation of proto-oncogenes. Cellular ...
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Is there a karyotype database for human cell lines?

I'm looking for the karyotype of a specific cell line. Some karyotype are well known, such as HeLa or some carcinomas, but some are very hard to find, such as LG2 (B cells). Is there a database ...
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Trypsin and cell culture

I am doing an experiment where I have treat the cells with a drug and calculate their counts. I would like to know if is bad to trypsinize the cells in consecutive days i.e. twice within 48 hours. How ...
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Why do some protocols require prewarming a liquid medium before inoculating?

For example, in this protocol for E. coli competent cell preparation, it says: Plate 10 uL E. coli BL21(DE3) cells on a LB-agar plate; incubate overnight (12 hours). Prepare 500 mL SOB medium ...
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41 views

How to grow fibroblast colonies from single cells?

I need to establish a cell line starting from single cells seeded on a 96 wells plate by a FACS sorter. The cells I am using are human fibroblasts RPE-1 cultured in F12 medium supplemented with 10% ...
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41 views

Why do epithelial cells arrest in response to serum?

Primary epithelial cells, for example human mammary epithelium, fail to proliferate (arrest) in serum-containing medium. Therefore, a common growth medium for epithelium contains pituitary extract ...
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80 views

Do I need to serially dilute E. coli cultures for optical density measures?

I am expected to track cell growth by measuring the incubating culture's cell concentration every 30 minutes or so. So my questions are: Why do I need to do serial dilution (assuming that I do)? Why ...
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28 views

Can the Staph streak method be used for culturing Neisseria on blood agar?

What I learned in class: Normally chocolate agar is needed to grow Haemophilus and Neisseria. The Staph streak technique can be used for culturing Haemophilus on blood agar by allowing the Staph to ...
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55 views

Are stem cell lines cryogenically preserved or commercially available like HeLa cells are?

There are HeLa cells, which were taken from sick cancer patient and now is growing worldwide in different laboratories for experiments. HeLa cells can be kept because they are immortal and so they are ...
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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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43 views

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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107 views

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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21 views

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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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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55 views

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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80 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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27 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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40 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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155 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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110 views

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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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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296 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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340 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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75 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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141 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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775 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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217 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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81 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 ...
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599 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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147 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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429 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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161 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 ...
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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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84 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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57 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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58 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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227 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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189 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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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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635 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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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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158 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 ...