Culture Cells

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Questions related to Culture Cells

Peripheral mononuclear cell, protein release ?

Hi, I collect and isolate peripheral mononuclear cells, by ficcol gradient, culture the cells for 48H. I search a protocol, to lyse the cells, after the incubation period, and to collect all...

03 March 2019 4,226 1 View

Does anyone know whether I can store a fresh biopsy in culture media at 4°C for 12 hours before processing it?

My project is to develop an in vitro cell culture model by dissociating a fresh salivary gland biopsy, culture the cells and perform viability assays using flow cytometry every 24 hours. My...

03 March 2019 8,716 3 View

Can someone help with human aortic smooth muscle cells (HASMC)?

Hello, I just started working with human aortic smooth muscle cells (HASMC) for the first time and experienced contamination problems. I purchased Lonza's human smooth muscle cell systems with...

18 February 2019 1,008 2 View

How could I empirically measure the diffusion coefficient of a lipophilic drug in a biodegradable matrix?

I have a hydrophobic drug with known molecular weight and drug solubility in an organic solvent. I want to print a biodegradable scaffold embedded with the drug and culture cells on it for as long...

02 February 2019 6,911 2 View

Can someone help with human aortic smooth muscle cells (HASMC)?

Hello, I just started working with human aortic smooth muscle cells (HASMC) for the first time and experienced contamination problems. I purchased Lonza's human smooth muscle cell systems with...

02 February 2019 6,076 2 View

Why are our mammalian cells rounding up during culture?

Our labs mammalian cells - A375, and HEK293- seem to be unhappy and are rounding up during culturing. I have checked mycoplasma contamination using a PCR kit, that turned back negative. I have...

28 January 2019 3,232 7 View

Which agent could I use to basify the cytosolic pH of cultured cells?

I do not want to change my culture media, so I look for a drug able to basify the cytosolic pH. The objective is to reverse the effect of an acidifying drug, so it would have to be some reagent...

11 January 2019 646 0 View

For PK15 and St cells cultured in biocreactor, the percentage of dissolved oxygen (for example, 40%) is calculated by pure oxygen or air?

When we culture cells in bioreactor, we need to control the percentage of dissolved oxygen. This percentage of dissolved oxygen is dependent on pure oxygen or air? Moreover, what should be pumped...

08 January 2019 2,821 3 View

Is there a CD4 cell line that can be used to track Th1 vs. Th2 population shifts?

I'm tying to design an experiment wherein a starter culture of cells is treated/challenged to trigger the release of cytokines into their media. This media would then be collected and added to a...

04 January 2019 1,471 1 View

Is there a standard method for isolating plasma membrane lipids from animal tissue?

I am working with sea cucumbers, and i am looking for a method for isolating plasma membrane lipids from a tissue. There is quite a bit of literature on analyzing plasma membrane proteome of...

01 January 2019 2,872 1 View

Do dendritic cells need to be cultured in cell binding plates to mature?

I am optimizing a maturation protocol for iPSC-DCs. The cells adhere strongly to the plates which makes it hard to harvest the cells. I was wondering if the cells need to be attached to a surface...

01 January 2019 9,795 0 View

How can be used the Embryonated eggs for the viruses isolation and identification?

Virus Isolation (Indirect Examination) : A. Detection of virus growth in cell cultures 1- Cytopathic effects (CPE) – morphological changes in cultured cells, seen under microscope, characteristic...

01 January 2019 7,638 0 View

LPS work better in normal conc. of FBS or reduced?

Dear experts, During in-vitro culture of cells, LPS works better in normal conc. of FBS or reduce in culture medium? usually I use 10% FBS. Either I use this conc. of FBS or reduce it for...

01 January 2019 9,128 2 View

Is there a CD4 cell line that can be used to track Th1 vs. Th2 population shifts?

I'm tying to design an experiment wherein a starter culture of cells is treated/challenged to trigger the release of cytokines into their media. This media would then be collected and added to a...

01 January 2019 5,826 1 View

Which conditions expansion hybridoma culture cell?

I would like to know which conditions of size and volume (Flasks T) for expansion/dead hybridomas cells?

01 January 2019 2,366 0 View

Why U937 cells are keep on detaching after PMA treatment ?

Hello everyone, I am treating U937 cells with PMA at 100nM concentration for 24 hr and almost 90% of cells get attached. Then I am culturing these cells with 10% FBS media for the next 24 hr....

01 January 2019 9,773 2 View

Statistical Analyses of Trypan Blue & Cell TitierGLO assays?

I'm developing an assay to determine sensitivity to a hypomethylation chemotherapy agent. Preliminary cytotoxicity assays were conducted to determine dosages that yield the optimum viable...

13 December 2018 2,106 3 View

Can anyone suggest an expression vector that is best to produce PIP2 positive vesicles in culture cells?

I am attempting to produce intracellular PI(4,5)P2 coated vesicles by introducing an expression vector for a constitutive active form of Arf6(Q67L). After that, I would like to observe large...

12 December 2018 3,068 0 View

Starvation-induced autophagy: friend or foe?

Autophagy induction in response to starvation may be a protective mechanism

12 December 2018 1,835 5 View

Problems with 3T3-L1 Adipocytes?

Hi there, I am new working with 3T3-L1 Adipocytes I am facing some issues with these cells. Can anyone suggest me how to overcome this? I purchased 3T3-L1 cells from ATCC and cultured these cells...

12 December 2018 3,435 10 View

Have anybody faced repeated contamination of AGS cell line?

Hi I have AGS cell lines in culture for our experiments. Recently, I started facing fungal/ yeast contamination of the AGS cell line in culture continuously, once they are 90% confluent. The...

12 December 2018 1,587 11 View

Are there any alternatives to measure cell death/apoptosis besides lactate dehydrogenase assay?

The LDH assay measures cytotoxicity. However, it requires culturing cells in media with less than or equal to 1% FBS. Are there any alternatives?

12 December 2018 3,926 3 View

Problems with 3T3-L1 Adipocytes?

Hi there, I am new working with 3T3-L1 Adipocytes I am facing some issues with these cells. Can anyone suggest me how to overcome this? I purchased 3T3-L1 cells from ATCC and cultured these cells...

05 December 2018 689 10 View

Considerations in using SH-SY5Y cells for toxicology studies; which population of cells do we consider for assays- floating or attached or both?

In our lab, we are trying to understand the mechanisms by which certain xenobiotic compounds induce neurotoxicity using SH-SY5Y cell line as a in vitro model. As of now, we are encountering a...

04 December 2018 6,291 1 View

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