Questions tagged [cell-culture]

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

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How to prevent HL-60 cell contamination?

I have thawed Hl-60 cells from liquid nitrogen chamber two times in RPMI 1640 media (containing 10% FBS, L glutamin, penicillin/streptomycin). In the first day the cells remain okay. But when ...
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Will dead cells in culture media have any significant impact on Live cells during incubation?

I am generating stable cell lines and have something like 25+ culture flasks to take care of. Ideally I can let dead cells sit in the media with the live adherent without rushing to change media ...
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Where am I going wrong in a Reactive Oxygen Species Assay done in N9 cells?

I have followed the modified protocol that was given to me by my senior (who was using for his RAW 264.7 cells - Reference article: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3308605/ ) for ...
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547 views

“producer cell line” vs. “host cell line” in biopharmacology

I'm translating a text that describes the creation and testing of a cell line that produces a drug (a protein) and also procedures for creation and maintenance of cell banks. Example sentence: ...
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1k views

Rescue cell lines function

I'm reading in a paper in which the authors have made three cell lines to study the function of a gene of interest (Mbd3): The one is knocked out for the gene of interest [Mbd3 -/-], the other is ...
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1answer
95 views

Measuring optical density with/without re-suspending cells

When measuring absorbance using a plate reader, is it necessary (or better) to resuspend the culture if the plate has been sitting still for 1-2 days? While this could be a good physics question, I ...
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69 views

Challenges of Industrial-Scale Fermentation

I was reading the wikipedia article on industrial fermentation, and it mentions that It is well established in the field of industrial microbiology that what works well at the laboratory scale may ...
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1answer
4k views

Purpose of dilution streaking or streak purification

In dilution streaking a drop of liquid culture is collected with an inoculating loop and then streaked across the agar plate surface. At the end of the streaking the number of organisms clinging to ...
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1answer
7k views

Is there a difference between “cell line” and “cell strain”?

I'm translating one Russian text where a cell line is transfected to produce a drug. I wonder if I can sometimes use "cell strain" or simply "strain" instead of "cell line" or instead of "producer" (...
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90 views

How do neuroblastoma cells divide? Is it via mitosis?

My name is Daniel and I'm 17 years old. I am currently designing a study to investigate compounds that potentially increase the replication of neural cells. I will be using neuroblastoma cells and ...
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1answer
370 views

How far is stem cell research from being to generate a new organ?

How far are we from being able to use stem cell research to generate a sperm, a heart, a liver, and a kidney? In stem cell research, my understanding is that you must start off with some cell to ...
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87 views

Did the Hela cells leave John Hopkins

Did John Hopkins let people outside of the university use the Hela cell-line that was isolated from an immortalised cervical cancer?
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876 views

Parafilm vs Foil for Retaining Plate Moisture while Allowing Gas Exchange

We often plate Streptomyces aureofaciens and grow them in a 30°C room. A post-doc I work with has always loosely wrapped the agar plates in aluminum foil to retain moisture and prevent cracking of the ...
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Effective Mycoplasma Elimination from Primary Human Cultures

Obviously the best way to avoid mycoplasma contamination is to avoid it in the first place. In our case, however, it is not possible to avoid. We are culturing viruses and tissues out of human nasal ...
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2answers
862 views

What kind of detergent should I use in cell culture laboratory for cleaning purposes?

I work on establish a cell culture laboratory in the company where I work. All equipment are new and I want to clean lab before I start to work with cells. Is there any special detergent for cleaning ...
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196 views

why selecting cells after gene editing with CRISPR Cas9?

I am new to the CRISPR Cas9 genome editing system and I have the above basic question. Another way to think about this: what would happen if after cells are transfected, the cultures are left to grow ...
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1answer
368 views

Do bacteria grow on pure dry glucose?

I've accidentally touched pure glucose with my bare hands (fingers to be specific), which was intended for cell-culture. I'm worried that bacteria from my skin may start to grow on the glucose and ...
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298 views

Respiration of an animal cell media

I want to do respiration process of an animal cell media in laboratory environment. I don't entirely sure how to supply it with Oxygen to the process because it is a gas, but not a liquid.
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157 views

Can you replace glucose with glycerol in cell media?

In order to feed an animal cell in process called Respiration, can I replace Glucose with Glycerol? The Equation bellow: Glycerol + Oxygen -> Water + Carbon Oxide
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1k views

Confluency or confluence, which term is correct to describe the % area covered by cells?

I noticed that both of them are used in many scientific papers. Are these two terms, or can they be used interchangeably?
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how to make plasma cells adhere to the bottom of a microplate?

I am isolating single plasma cells by FACS sorting into 384-well plates, with the intent to assay the supernatant and clone H/L chains from positive wells. The efficiency of the PCR is however low, ...
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1answer
16k views

Formula for number of divisions required to form x number of cells (Mitosis and Meiosis)

In case of Mitotic Division, we say that if the total number of cells formed by mitotic divisions are $x$, then counting from the first cell onwards, we get $x-1$ divisions ($N$) $N = x-1$ This is ...
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1answer
46 views

How to induce a steady increase in cyctoplasmic [Ca2+] in HEK293T cells? And is there any simple marker/method to confirm it?

We are trying to find out whether increase or decrease of cytoplasmic calcium concentration affects the interaction of two proteins, using co-IP in HEK293T cells as a readout. In different forums and ...
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1answer
302 views

Soft Agar Substitute

I've got petri dishes with a base agar layer ready for E. coli culturing. However, I do not have soft agar in which to dilute the E. coli before pouring it on to the plate. Any suggestions on what ...
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1answer
346 views

What are mouse LS cells?

I have seen the term "mouse LS cell" used in several papers now, but I haven't found an explanation for the origin of these cells (for example, this paper). What are mouse LS cells? What is their ...
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Why lactate inhibits growth (or enhances death rate)?

Extracellular lactate tends to inhibit cellular growth or enhance cell death. This happens in the vicinity of tumors and in cell cultures. See for example this reference: Ozturk, Sadettin S., Mark R....
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60 views

Does a specific blood group enhance the Plasmodium growth?

I am maintaining Plasmodium falciparum cultures for past 6 months. For the blood culture, usually we lab members take turns and donate blood for the culture. I observed that the parasite's normal ...
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1answer
101 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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86 views

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
72 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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768 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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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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614 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 ...
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1answer
38 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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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 ...
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104 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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561 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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871 views

Is there a karyotype database for human cell lines?

I'm looking for the karyotype of a specific cell line. Some karyotypes 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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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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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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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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2k 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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1answer
674 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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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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45 views

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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1answer
2k 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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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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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 ...