Questions tagged [cell-culture]

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

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1answer
198 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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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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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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147 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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59 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
24 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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21 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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97 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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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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189 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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1answer
34 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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How do I interpret this graph regarding introduced genes and virus-infected cells?

This graph appeared in a practice test for the MCAT. I am trying to interpret it, but it confuses me a bit. On the x-axis we have some introduced genes, and on the y-axis we have % of cells infected. ...
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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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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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119 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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20 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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64 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 ...
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Does trypsin strip flask coating?

Mammalian cell/tissue cultures sometimes require flasks coated with proteins. My uneducated guess is that these proteins mimic the ECM, perhaps the basal lamina, so finicky contact-dependent cells can ...
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1answer
51 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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How long does antibiotic-dosed LB maintain good selection?

Various people in our lab will prepare a liter or so of LB, add kanamycin to 25-37 mg/L for selection, and store it at 4 °C for minipreps or other small cultures (where dosing straight LB with a 1000X ...
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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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92 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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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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36 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 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?
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35 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
55 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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2answers
300 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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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 ...
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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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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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2answers
358 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
416 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?
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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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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)
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33 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 ...
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173 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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56 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 ...
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2answers
2k views

How much time do INS1-E and MIN6 cells need after splitting?

I am currently doing an experiment on cells to test the internalization of a protein. Normally, I seeded my cells the day before the incubation. This worked well for Hela, CHL or PANC1 cells. However, ...
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258 views

Which human cell lines do not express the GLP-1 receptor?

I need a human cell line that does not express the GLP-1 (glucagon like peptide-1) receptor. I'm working with HeLa cells, do those express the GLP-1 receptor? Which other cell lines exist that don't ...
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2answers
84 views

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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1k views

Are HeLa cells edible?

I'm curious if HeLa cells are intrinsically poisonous or dangerous to ingest. My understanding is that some of the contamination in HeLa cells such as HPVs are not readily expressed. I have no plans ...
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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 ...
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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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265 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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7answers
6k views

Pipetting damage to cells

I'm curious how much damage is potentially inflicted by shear stress by pipetting. I know that syringes used for stem cell injection can cause a lot of damage. However, to what extent does this happen ...
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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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579 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 ...