Questions tagged [cell-culture]

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

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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 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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1answer
106 views

Microinjection intestinal organoids

I am attempting to microinject small intestinal organoids, using the Eppendorf FemtoJet 4i microinjector and Femtotip capillaries, and have come across several issues. This is a new set up in our lab, ...
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Low cost cell free expression system

I am a graduate student looking to incorporate some protein engineering into my training. I have been reading a lot about E coli lysate based systems, but they still require 3rd party additives like ...
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290 views

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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459 views

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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37 views

Pharmacokinetics and cell-membrane permeability of Adenosine triphosphate disodium hydrate

【My Question】 (1) Please tell me the pharmacokinetics of this ATP (or Adenosine triphosphate disodium hydrate) when it is administered orally or intravenously. In particular, Does this material has ...
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1k views

What is the difference between enrichment media and selective media?

Enrichment media These media are used to suppress commensal bacteria while allowing the pathogen to remain viable and grow. [Source: Ananthnarayan and Paniker's textbook of Microbiology.] ...
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339 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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37 views

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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79 views

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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106 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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187 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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27 views

What culturing process / experiment needs a “grid” petri dish (one with a vertical division)

You can buy petri dishes with separations in them like this: I have studied biology and biochemistry and never come across one of these before. What cell culturing / experiment would this be used in?...
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22 views

Learning industry skills for virology?

I am trying to learn more about virology for a tentative job I could be taking soon, as a quality control person in virology. However, I have not taken a formal virology course, nor any course work ...
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21 views

Beyond anabolism, what are factors that restrict growth rate?

Beyond obvious constraints in the case of nutrient limitation, what can explain the relatively slow growth rate of cells in rich media? Lab strains of E. coli grow really, really fast. The cell ...
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14 views

Can you gain valid toxicological data about a detergent in cell cultures?

I just looked at a paper that in one assay tested a variety of chemicals and chemical mixtures against human cell cultures. They exposed HEK 293 cell cultures for 24 hours to the various chemicals. ...
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37 views

Isolation of Intact Granules from Mast Cells

How to isolate intact granules from mast cells without using sucrose and percoll?
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What is(are) the mechanism(s) that stop cells from fusing in vivo?

I just learned about the phenomenon of 'cell fusion' in which two diploid somatic cells can combine into some aneuploid cell in vitro and proceed to proliferate in culture. Apparently this can even ...
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45 views

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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34 views

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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19 views

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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209 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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72 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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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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941 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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108 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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89 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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2k views

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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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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7 views

What is the typical range of volumetric productivity and cell density in aerobic fermentor bioreactors for microes?

Im doing research on biomass production. Its hard to find data for this question. I need citations to calculate the production rate of microbial biomass in bioreactors (e.g aerated fermentator). ...
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19 views

Continuous cultivation cell growth - help with understanding calculation

In the solution (blue text) what is the term µCxVL? And also could you help me understand the meaning behind Cx(µ-0,10), what does it actually mean? Thank you!
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Why do people use J2 viruses for macrophage immortalization?

I've been trying to figure out how to go about immortalizing some primary alveolar macrophages harvested from mice. What I am seeing in the literature is that it seems like people have been using a ...
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22 views

Why don't materials and methods describe culture media in volume?

After reading several materials and methods sections of cell culture experiments, I have not seen the recipe for culture media ever described in terms of volume. Human adipose tissue was obtained ...
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14 views

Length of cell cycle dependancy on nutrients, pressure, temperature or other factors

How is the duration of cell cycle influenced by the concentration in nutrients, mechanical constraint, temperature or other factors ? And can it change strongly ? Meaning there are 2 possibilities, ...
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1answer
46 views

Approximate Cell Doubling Time?

If I know it takes 12 min to occur in mitotic fibroblast cells and I count there are 32 dividing cells in the microscope slide out of a total population of 32034 cells, how do I calculate the ...
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1answer
195 views

Inoculating agar plates with microbes suspended in sterilised water?

We were having a problem with our experiment (culturing microbes on homemade nutri agar) so we decided to change our set-up. It already cost us quite some money. So, we saw this procedure. Can anyone ...
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137 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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198 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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641 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
233 views

What is the recommended culture volume for HeLa cells and seeding number for 384 well plate wells?

For 24 well plate wells, thermofischer recommends to seed 0.05E6 cells with 0.5 to 1 ml of medium. What are recommended numbers for a 384 well plate well?