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Questions tagged [cell-culture]

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

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0answers
788 views

Two different sized colonies from the same species of bacteria. What does it mean? [closed]

I have got two different sized colonies in a plate of Salmonella Paratyphi, identified by biochemical methods. What does two different sized colonies mean? What kind of question can I investigate in ...
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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 ...
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1answer
105 views

Gender of BEAS-2B cell culture

I recognize some interesting patterns when analysing DNA methylation of BEAS-2B cells. Does anybody know, if the cell culture is female or male? I already asked the customer support of Sigma Aldrich ...
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1answer
597 views

How long can a human neuron live outside the body in a controlled environment?

Have there been any experiments that have kept neurons alive (stationary), without preserving methods such as freezing? If yes, then how long were the cells kept alive for?
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UACC-462 cell culture

UACC-462 is a pancreatic cell line established in the University of Arizona Cancer Center (UACC). The cells are available from ATCC and there is the instruction for culturing UACC-462. M-41 medium. ...
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1answer
74 views

Is it possible to grow E. coli K12 with algal amino acids as the sole carbon source?

Instead of glucose or glycerol, is it possible for E coli to grow on an amino acid mixture as the sole carbon source and, if so, what should the concentration be?
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1answer
43 views

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 ...
2
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1answer
2k views

Staining cells for FACS at 4 degrees or ambient temperature

I'm sorting a 293 derived line. One thing that is worrisome for me is cell viability after the sort. Usually I have been staining and washing at room temperature (on the benchtop) on a nutator. I have ...
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662 views

Jurkat single cell clonal expansion

I'm hoping to do a clonal expansion of Jurkat, expressing my construct which is currently transduced virally. From the FACS data I currently have roughly 30-40% expression in my culture but I need it ...
11
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1answer
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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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1answer
39 views

Testing CO₂ for cell culture

I'm am refurbishing some old CO2 incubators. I would like to test the precision of the CO2 sensor. There is a sample port on this incubator. What sorts of methods are commonly used to measure CO2 in ...
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107 views

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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175 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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614 views

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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2answers
1k 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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3answers
2k views

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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1answer
2k views

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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4answers
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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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1answer
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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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1answer
717 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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1answer
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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 ...
3
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1answer
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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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1answer
1k 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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1answer
149 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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1answer
238 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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1answer
489 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 ...
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1answer
202 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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1answer
83 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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1answer
94 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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3answers
972 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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1answer
55 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?
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1answer
270 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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1answer
105 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 ...
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0answers
861 views

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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3answers
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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?
4
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1answer
219 views

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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189 views

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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1answer
247 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?
3
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1answer
119 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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1answer
3k 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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54 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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1answer
238 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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1answer
4k 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, ...
4
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1answer
305 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 ...
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1answer
3k 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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2answers
3k 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 ...
2
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1answer
397 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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2k 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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2answers
637 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
4k 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 ...