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Questions tagged [cell-culture]

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

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22 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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20 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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12 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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19 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
105 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
34 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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1answer
39 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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918 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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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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1answer
170 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?
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1answer
32 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
42 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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26 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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21 views

Micro-propagation of Moss in Agar (sterilization?)

I'm having trouble sterilizing my moss sample for micro-propagation. Moss is basically a sponge and it has a massive surface area. I tried Isopropyl alcohol figuring it had a very high probability of ...
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33 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
58 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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13 views

Need to learn about basics of eukaryotic cell culture and transfection

Can someone please suggest books/ online courses/ other resources to learn about cell culture and transfection theory as well as techniques? I've been reading research papers and everyone uses methods ...
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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
40 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)
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1answer
85 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
31 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
26 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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1answer
126 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
54 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
229 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
349 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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1answer
146 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
290 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
47 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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1answer
169 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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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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0answers
42 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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0answers
242 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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42 views

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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32 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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0answers
2k views

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
202 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
528 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
63 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
433 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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123 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
51 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
526 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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0answers
18 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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200 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
42 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
118 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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0answers
305 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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0answers
60 views

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 ...