Questions tagged [cell-culture]

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

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Why E.Coli get bigger if they replicate faster?

When E. Coli divide faster, they tend to have larger volumes. If the same cells are grown in a poorer medium, they'll have longer replication cycles and shrink. What is the explanation of this ...
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Are there any techniques for manufacturing exosomes in the lab?

I'm interested in the idea of using exosomes as an alternative transfection agent / delivery mechanism for gene editing applications (CRISPR/cas9, etc). Information on the matter is sparse.. Does ...
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How long can one go with necrosis?

In general, when a limb dies, how long does it take before it falls off? Can you go years with a black and useless limb, or does it just.. fall off right away?
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Can anyone expolain this? Cell culture vials have strange hair-like condensation patterns

What is the hair-like polymer that condenses on the caps of the plastic cell vial? Inside vial are cells in a mixture of FBS and DMSO. The vials were frozen slowly in a special isopropanol-filled ...
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How long can you store tissue on 4°C before B-cell extraction and culture?

We receive tissue regularly and extract B-cells for culturing, studying differentiation and survival with diffenet stimulation conditions. However, often the tissue arrives late afternoon and might be ...
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Why is a DMSO-only Control Important? [closed]

Why is it necessary to have a DMSO alone control when DMSO has been used as the solvent (vehicle) in the treated conditions? And, in this case, why is the viability of the untreated condition lower ...
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1answer
336 views

Flow Cytometry to learn about Cell cycle

I am posting my Flow Cytometry data here again because I realize that there are many people who are willing to help me, and I can learn a lot from you guys. In this cell culture Lab, we use Cell U-...
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1answer
692 views

Flow cytometry data Analysis [closed]

everyone. Since I am new in the flow cytometry technique, I am not really sure how to analyze the result. We use cell U-937 Lymphoblast.All Cells Could Anyone help to explain me this one? Thanks ...
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1answer
69 views

How is an RT-PCR representative of the RNA contents in a cell?

For example, we first collect all the RNA contents from a mammalian cell line. Say, for instance, we collect 80 uL of RNA with a concentration of 150 ng/uL. Next, to make cDNA, we take 1,000 ng of the ...
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1answer
518 views

Trypsin and Cell Surface Receptors

Can adding Trypsin to mammalian cells affect the expression of or hydrolyze cell surface receptors?
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148 views

Pelleting bacterial cells using centrifugation and getting proteins in the supernatant

Do significant amount of proteins from cells find their way in supernatant during pelleting? If the answer is yes then I see two ways in which protein can be in the supernatant . 1) Through lysis ...
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1answer
59 views

Estimation of cell count of E.coli culture

Can I get some help with an estimation of a cell count in a culture of E.coli cells if we assume we begin with $10^6$ cells, that they divide every 30 minutes and they are incubated for 12 hours? I ...
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1answer
653 views

Mass production of bacteria

Is there a way to mass produce a certain species of bacteria? Answers I am looking for should not involve a Petri dish: I am only interested in protocols involving, perhaps, more efficient mediums to ...
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In Vitro Meat Initial Production

I've gotten interested in the science behind in vitro meat, and I was wondering what would be necessary to create it. Would you initially need a primary culture, or an immortalized cell line? And how ...
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212 views

Why are my assay absorbance values very low?

I am culturing SH-SY5Y cells in 96-well plates at a seeding density of $1.2 \times 10^5$ cells/mL. I differentiate the cells with retinoic acid for 6 days before giving them treatments. The cells look ...
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1answer
47 views

Why would you plate cells at different densities in a colormetric assay?

The aim of my experiment is to see if a kinase inhibitor reduces cancer cell viability. I am using 2 different cell densities 50,000 cells/well and 100,000 cells/well and different doses of the kinase ...
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3answers
141 views

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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347 views

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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1answer
620 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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1answer
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
100 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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73 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
5k 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
8k 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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95 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
379 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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1answer
88 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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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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113 views

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
1k 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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199 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
385 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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1answer
354 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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1answer
166 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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1answer
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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18k 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
49 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
339 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
379 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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1answer
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
106 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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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
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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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789 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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682 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
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 ...