Skip to main content

Questions tagged [cell-culture]

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

Filter by
Sorted by
Tagged with
6 votes
1 answer
58 views

dilutions in bacterial culture

In overnight bacterial culture, why do we put bacteria in a small flask to culture before moving it to a larger flask? Why not just put the sample in a large flask to begin with? This is in reference ...
1 vote
1 answer
53 views

Image Dataset for Cardiac cells for Mus musculus?

I'm working on developing a transfer learning model to detect cells in a culture using microscopic images. for that i require data set of images having microscopic images of various cell types. Is ...
1 vote
0 answers
23 views

What is the largest culturable bacterium?

E. coli is easy to grow in the lab. It has a volume around $0.6 \mu m^3$. Source T. namibiensis can be much larger, with a volume on the order of ${10^6}\mu m^3$. Source However, T. namibiensis is not ...
0 votes
1 answer
80 views

Is this contamination, debris?

I have been culturing SVG cell lines. This line was obtained as a live culture from another institute and was delivered by mail. After subculturing , I saw some particles, and I do not know what they ...
0 votes
1 answer
29 views

Plant tissue cultures without Phytohormones?

Is it possible to do tissue cultures without using phytohormones like BAP or NAA. So to just use for instance MS-Media. Online, I just found that every plant is possible to culture with the right ...
1 vote
1 answer
22 views

Cell viability assay: Problems with MTT assay in the solubilization step

I am testing two compounds against 3 cell lines to determine cell viability. Two lines grow in Eagle media, one in DMEM (+10 Percent bovine serum) which contain phenyl red indicator. I prepared 3 ...
0 votes
1 answer
59 views

Why does my cell subculture have these black dots like appearance?

I tried to subculture cells(hek293 cells) for the first time for a Cell Biology experiment, and here are the steps I followed:- Cell subculture (*the media is a mix of 10:1:0.1 of DMEM:FBS:Pen/Strep) ...
0 votes
0 answers
17 views

For Cytotox studies of anticancer compounds: What cell line can I use to check if my compounds will also damage healthy cells?

I am about to perform cytotox studies on cells with my synthesized compounds. I want to test my structures against MCF7 cells (breast cancer) as well as a melanoma cell line. MCF7 should be vunerable ...
1 vote
1 answer
52 views

How much effort is it to establish a cytotox assay for cancer cell lines against a small number of possible compounds?

I am currently testing a series (5-10) of small molecule compounds against an enzyme that are intended as inhibitors. This enzyme is meaningful for cell proliferation. Until now, nothing was active ...
0 votes
1 answer
52 views

Why are there error bars for zero concentration in some cell viability graphs, if they are assumed to be 100%?

Take these for example: As far as I know, the formula for cell viability is % Viability = (Treatment OD - Blank OD) / (Control OD - Blank OD) where each OD is the average of three wells (technical ...
0 votes
0 answers
31 views

How to keep primary human cancer cells alive in vitro and derive a cell line?

I am studying a human cohort of patients treated with neoadjuvant immunotherapy before surgery, of whom we have pre- and post-treatment biopsies as well as the resection. In one patient's resection, ...
4 votes
1 answer
910 views

Bacterial cells disappearing from culture with time

While growing my bacterial culture in liquid media for protein expression, I have been observing that the bacterial cells (E. coli) grow well for some till OD600 reaches ~0.4-0.6, then suddenly ...
0 votes
0 answers
18 views

Cell culture disasosiation

I am doing to passaging cancer cell ( Dld-1 Cell line). Trypsination was conducted with 0.25 trypsin-edta 1 ml to t-flask 25 in 2 min. Then I add 6 ml complete media (have 10% FBS for inactivated ...
1 vote
0 answers
22 views

MTT assay data pre-processing

I was handed MTT assay data, consisting of drug doses and raw OD scores. The drug treatments were performed three separate times, each followed by MTT assay. I applied some non-linear mixed-level dose-...
1 vote
1 answer
68 views

Can a non-pure culture be used for reliable rapid phenotypic diagnostic tests?

Can a non-pure culture be used for reliable rapid phenotypic diagnostic tests (e.g. catalase, oxidase & gram stain)? "Slide Test Method Obtain a pure culture of the organism to be tested. ...
0 votes
1 answer
37 views

Induction of IFN-beta in HEK293T

I'm trying to increase expression of a protein we're attempting to study, UBL7, supposedly unregulated by Type I Interferon and particularly IFN-beta. I've tried treating HEK293T cells (~60% ...
0 votes
1 answer
47 views

Where can I find a video of human cells reacting to cold water vs warm?

I have the following lists: ...
1 vote
0 answers
25 views

What are common culture temperatures?

In literature, authors sometimes refer to 37°C as a common culturing temperature. As I understand it, those are temperatures that are conventionally used for incubators in laboratories. For mesophilic ...
0 votes
1 answer
101 views

What kind of bubble this?

This is the MARC-145 I cultivated. Recently, many bubbles appeared in the cells. I don’t know why.
0 votes
0 answers
28 views

Cell culture contamination

I've been culturing my cells over 2 weeks and just yesterday these things popped up in my flask , I'm not sure if they're contamination or not.uesterday they looked white and today I found one of them ...
2 votes
1 answer
54 views

Would BMP7 introduced to a Petri dish with cartilage cause chondrocytes to replicate?

Chondrocytes are the cells that make up cartilage. As I understand it, the best growth factor that signals replication of chondrocytes is Bone morphogenetic protein 7. If supplied with a sufficient ...
-2 votes
1 answer
61 views

Need help identifying this microbe

Sorry for the bad image, we don't have a camera hooked up to our culture room microscope yet. But Has anyone seen these kind of (what I'm assuming are) contaminating bacteria before? We see these from ...
0 votes
1 answer
270 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 ...
0 votes
2 answers
197 views

Is there a way to relate the OD to cells/mL without doing a standard growth curve?

I am helping out with an experiment with multiple bacterial cultures (>20 species). I need to a very specific concentration (cells/mL or CFUs*/mL) for this purpose. Unfortunately it appears that ...
0 votes
1 answer
45 views

Is somatic fusion/hybridization off-limits to DIY-ers? ELSE: how can an infertile plant's characteristics be salvaged to a fertile one?

I was amazed to discover plant tissue culture could be performed relatively easily at home. I have been wondering, what's the likelihood of a DIY-er being able to setup and learn a crude and basic lab ...
0 votes
0 answers
87 views

Why is my spirulina biomass getting destroyed in my new photobioreactor?

I have a 500L flat plate photobioreactor in my backyard. Whenever I add my culture to the PBR, after 3-4 days of growth, the culture becomes transparent and my biomass is wasted. I add the stock at a ...
1 vote
0 answers
49 views

Does decellularization with sodium lauryl sulfate break down enzymes?

I am currently looking into decellularization with sodium lauryl sulfate since it is effective and cheap. My goal is to break down the cells of a plant leaf and release the chloroplasts to be viewed ...
1 vote
0 answers
19 views

Endospores contact with surfaces

I am currently studying endospores-forming bacteria. To be specific B.subtilis. I noticed that the formed spores have strong contact "grip" with the surface it is forming on. For example, ...
0 votes
0 answers
29 views

Single-cell subcloning of BV-2 cells?

I am trying to knock-out genes in BV-2 cell line. However, the majority of protocols require cell subcloning and expanding. I tried to subcone these cells and grow clones in DMEM+10% heat-inactivated ...
0 votes
0 answers
59 views

What is the solubility of rotenone in ethanol?

I’m trying to kill SH-SY5Y cells with rotenone, but the rotenone does not kill. I saw an article where rotenone is soluble at 5mM in ethanol. I dissolved 1.7 mg in 1 ml of ethanol to obtain a 1ml of ...
1 vote
1 answer
4k views

RNeasy Mini Kit low 260/230 ratio -- can I purify this RNA for further use?

I used Qiagen's RNeasy Mini Kit to isolate RNA from 5*10^5 C28/I2 (immortalized human chondrocytes). However, my yield is low (~25 ng/ul), but my 260/280 ratio is great (~2.3), and my 260/230 ratio is ...
3 votes
0 answers
77 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 ...
-1 votes
1 answer
144 views

Does centrifuging the bacteria culture removes bacteria leftovers?

Usually, we centrifuge bacteria culture to either replace the media, control its density (OD), ..etc. I know that when centrifuging the cells are pushed to the bottom while the media (solution) ...
1 vote
1 answer
180 views

Sabouraud dextrose agar breaking?

I recently started growing some Geotrichum Candidum (GC) on Sabouraud dextrose agar (SDA) beds. The SDA was poured into sterilized petri dishes. After setting the plates in a cooler (20 C) for a day, ...
0 votes
0 answers
1k views

What is the difference between BSA fraction V and the initial fraction?

I am preparing a supplemented culture medium and I need to use BSA. There is fraction V of BSA that we normally use for immunocytochemistry and there is another type named the initial fraction that is ...
0 votes
0 answers
781 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 ...
4 votes
1 answer
2k 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 ...
0 votes
0 answers
106 views

Fixing the cell with 4%PFA and staining with PI to observe cell cycle?

I am wondering whether we can replace 70% ethanol to fix and perm the cells with 4% PFA to observe the cell cycle. Do I understand correctly that 4% PFA can permeate the cells, thus the PI will ...
1 vote
1 answer
151 views

Appropriate cell lines to study depression

Short introduction I'm studying depression from a biochemical point of view. My interest lies in the study of protein biomarkers and I was wondering which cell lines may be appropriate for this ...
0 votes
0 answers
494 views

Virus culture in artificial/synthetic medium

I am looking for a publication/paper in a well a circulated magazine/journal/government study on growing virus culture in synthetic or artificial medium. I have found this link on SARS Cov 2 being ...
0 votes
1 answer
106 views

What is the meaning of harboring cryptic in below sentence?

Tissue culture became more widely used at this time because of the introduction of antibiotics, which facilitated long-term cell line propagation, although many people were already warning against ...
9 votes
1 answer
199 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, ...
1 vote
0 answers
25 views

How do I cover Cellstrainers?

I‘m doing some experiments for my bachelors thesis and I‘m using some Cell strainers by Roth for it (https://www.carlroth.com/com/en/accessories/cell-strainer-easystrainer™/p/cly9.1). My problem is ...
1 vote
1 answer
257 views

How to create long term stock for Bacillus subtilis?

I am trying to create a long-term stock for Bacillus subtilis (Bacterial glycerol stocks). I have looked online for guides and step-by-step tutorials and I found that they don't say where the sample ...
1 vote
2 answers
3k 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 ...
2 votes
1 answer
99 views

Decreasing signals in assay measurements

I'm working with a calcium assay to study the effects of different virulence factors. The assay works, but from day to day the signals of cell lysis go down. Unfortunately, I haven't found an ...
0 votes
0 answers
47 views

CML (Chronic Myeloid Leukemia) and blast cell percentage

I'm trying to understand the oncogenesis of CML. I have a question about CFC cells and blast cells. Are those the same? I know that in the chronic phase of CML there is a blast percentage of 1-10% of ...
4 votes
2 answers
1k views

Why does vacuum sealing of foods prevent spoilage from anaerobic bacteria?

I have noted that many purchased food items in containers only require refrigeration after they have been opened, thus eliminating the vacuum seal. Air contains bacteria and fungus spores that enter ...
0 votes
0 answers
38 views

QA/monitoring in recombinant vaccine manufacturing

On a16z[0] podcast on vaccines[1], an interesting tidbit came up: vaccine manufacturing in cell cultures is expected to hit a major challenge in terms of quality control due to unpredictable behaviour ...
3 votes
1 answer
84 views

Can a cell culture insert plate be used with a vacuum manifold?

There exists 96 well plate setups for use with vacuum manifolds. Is it possible to use 6/12/24 plate inserts with the same setup?

1
2 3 4 5