Questions tagged [cell-culture]
Cell culture is the process of growing and maintaining living cells in vitro.
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Is it safe to work with HeLa cells?
Hela cells are infected with HPV. So is it safe to work with them? What are the safety precautions?
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1answer
40 views
Selection marker for a cytoplasmic vector that works at low expression levels?
I need to express a selection marker on a plasmid in human cells (e.g. HEK293). However, I am using a cytoplasmic vector derived from a non-human virus with its own mechanism for transcriptional ...
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28 views
How can I propagate a community of anaerobic bacteria sampled from mud in a bog?
I'm doing research on anaerobic bacteria. In the fall I took mud samples from a bog and have been working with them but I didn't take enough. Now winter has rolled around (Im in Canada) and all the ...
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58 views
Can microbes become “overweight”?
Just out of some weird thoughts, can a microbe (any single-celled organisms, accepting answers for both prokaryotes and eukaryotes) ingest too much food (e.g. via absorbing too much involuntarily ...
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18 views
Using a magnetic stirrer as a makeshift shaking incubator?
I'm working in my high school's lab and need to culture bacteria in a broth culture. More specifically, I'm trying to harvest Bacillus endospores with the exhaustion method published in "Molecular ...
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1answer
32 views
Can primary tumor cells grow in 2D culture? [closed]
Is it possible to expand primary tumor cells in 2D culture? Are they adherent cells? Do you have any experience especially with culturing non-small cell lung cancer in 2D?
Thank you.
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25 views
What is the most durable electronic pipette?
We have a routine task of pipetting substance concentration series in triplicate on cell cultures under a laminar hood. To speed up the process, we use an electronic pipette in the repetitive ...
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0answers
17 views
Are controls needed for urine cultures?
If testing for bacteria (gram-negative) in a urine culture, are negative or positive cultures needed?
I could not find anything that says controls are needed unless results seem to be contaminated. ...
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18 views
Which culture media is suitable for cheek epithelial cells?
A month ago, I tried isolating cheek cells from saliva and the experiment was a success, however, I was wondering if it is possible to cultivate these cells in vitro. I'd say it's possible, but I wasn'...
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1answer
102 views
A biological system to measure time
First of all I am a physicist, so I apologise for my non-existent understanding of basic biology. However, I have what I think is an interesting question and this seems like a good place to ask it...
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26 views
Is it possible to grow a part of a plant independent of the rest?
Is it possible, using modern knowledge of biochemistry and synthetic biology, to grow a tomato without growing the rest of the tomato plant?
As an academic exercise, with full knowledge of the fact ...
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1answer
43 views
“Spontaneous Immortalization”
HaCaT keratinocytes are described as "spontaneously immortalized "(see paper below) due to aneuploidy or chromosomal alteration.
From the paper I get that this is a literal term, that the cells just ...
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9 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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22 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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40 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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0answers
41 views
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 ...
4
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1answer
173 views
Why is glycerol supplied/used as 87% solution?
While preparing glycerol stocks of my transformed bacterial cells (DH5alpha) the protocol I had to follow was that I would make one 50% glycerol stock (500µl culture, 500µl glycerol 50%) and another ...
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1answer
67 views
Why do many cells together survive better than one cell alone?
I know that one cell with no ECM is subject to anoikis. That's why in general one cell alone not in its environment is dying.
But are there advantages for a cell to live close to other cells even of ...
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1answer
110 views
Is my cell-culture contaminated with mold?
I am growing cell cultures at the moment, and I sometimes find 1-2 of these elongated, black structures in one of my 8 wells. Should I be concerned?
I have been growing these for about 5 days at ...
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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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0answers
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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0answers
30 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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1answer
55 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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23 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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0answers
24 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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0answers
22 views
How to take precise/low number of cells from cell lysate? [closed]
I would like to know that how can we take low or precise number of cells from a cell lysate? (without using any protein quantification assay).
Let say,
If I have a cell line having cell density 5x10^...
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1answer
122 views
Continuous (yeast) culture system satisfying multiple criteria
There are multiple ways yeast cells can be grown in liquid cultures.
Batch cultures are easy to prepare but cell density, medium composition and physiology change over time
Chemostats allow to ...
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1answer
195 views
Are cryovials really better than regular tubes for cryopreservation?
What is the rationale for using cryovials instead of regular, sterile centrifuge tubes for the storage of mammalian cells?
With a good aseptic technique and without touching the cap directly, there ...
2
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1answer
89 views
What does it mean to use DMSO as a dissolvant in biology experiemnts?
In many biology publications, I saw that dimethyl sulfoxide (DMSO) is used to dissolve drugs. However, the cell-based assays are still performed in water-based medium such as RPMI or PBS.
My ...
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0answers
2k 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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0answers
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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1answer
261 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?
2
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1answer
36 views
Growth mediums required for human cell cultures
So i would like to try a culture some of my keratinocytes just to see if i can do it. Ive read about a few different types of growth medium such as DMEM and RPMI but i am not sure about when to use ...
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1answer
81 views
Major effects on cells grown with 5% FBS supplemented media instead of 10% FBS?
I maintain a few cell lines in the laboratory (industrial R&D) that I use regularly for experiments. I realized today that the media i prepared for the week on monday was only supplemented with 50 ...
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0answers
30 views
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 ...
3
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0answers
38 views
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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1answer
141 views
Chunks of cells after trypsinization
Recently I've came across a problem after trypsinization of my stably transfected CHO-cells. When I add medium to rinse off the cells from the bottom of the flask, there are usually big chunks of ...
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1answer
59 views
Minimum hardware and knowledge to study effects on live cells [closed]
I'm very interested to study the effects (or reactions) of different forces (magnetic fields, electric fields, etc) on living tissues/cells.
I know I have to have a living cell, the force applied and ...
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1answer
65 views
What are the meaning the nomenclature in high cell density cultivation for the production of a protein? [closed]
I'd like to know the meaning of these terms in a HCDC studies of a modified strain.
Max specific yield (mg/g DCW)
Max volumetric product yield (g/l)
Specific product formation rate Qp (mg/g DCW/h)
5
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1answer
116 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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1answer
35 views
Is there a standard way of measuring culture nutrient levels in real time?
I am working on optimizing growth rates for a culture, and one thing I'm trying to monitor is nutrient consumption rates to pinpoint any bottlenecks, i.e. the culture goes through all the calcium in a ...
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2answers
27 views
Do PC-12 cells have to be stored in nitrogen?
My lab has been storing their PC 12 cells in the -80 freezer... upon arriving here and investigating a new project I see that it specifically stated to be stored in nitrogen vapor? Is this the ...
3
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1answer
198 views
Is it possible to make anaerobic media too reducing?
I've been struggling to grow an anaerobe (P. gingivalis), I take every precaution to keep the culture conditions free of oxygen, I was wondering if the conditions can be so reducing that they inhibit ...
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1answer
59 views
Tumour cell injection into a mice [closed]
I am going to inject mice tumour cells into mice to create them tumours and I am wondering if I need to have special precautions for that, even they are cells from mice tumour I am afraid what can ...
3
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2answers
399 views
What culture medium should we use for tap/drinking water bacteria?
We are a group of high school students from Manila who is performing an investigatory project on a topic that we chose which, at first glance, seemed simple, but turned out a bit complicated. We're ...
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1answer
519 views
Do dead cells always contain no nucleus?
If I examine a dead cell, can I be sure that it has not a nucleus?
And what about the other organelles?
2
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1answer
319 views
Mold identification
I poured some agar plates and tried inoculating them with mushroom spores (I don't think they germinated bc of low humidity), but noticed what looks like mold with crystals on the surface of the agar.
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2answers
423 views
293T safety, transfections with plasmid DNA
I am a little concerned with 293T cell lines, I have been working with this cell line for a few years with less frequency. However there were times that i extracted the proteins from these cell lines ...
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1answer
49 views
Yeast contamination with fungus patch
I am working on yeast mutant and I am observing their sporulation efficiency on agar plates. One of my plates got some dark green small fungus patches. I can't grow yeast again and dump this one with ...
