Culture Techniques

Methods of maintaining or growing biological materials in controlled laboratory conditions. These include the cultures of CELLS; TISSUES; organs;... | Contact experts in Culture Techniques to get answers

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Questions related to Culture Techniques

Measuring protein content of lung tissue for ACE assay?

I am measuring ACE activity in lung tissue and have homogenised it this way: Samples (~ 50 mg) homogenized in 300 μl ice-cold buffer, then centrifuged at 16400 g for 10 min at 4°C. Supernatant...

05 May 2016 9,974 1 View

Cell culture contamination with suspension cells?

I am culturing primary human brain stem cells which grow as neurospheres but I seem to constantly get what looks like bacterial contaminations and cannot seem to work out why. I have had 4 years...

05 May 2016 9,091 9 View

Can anyone suggest the culturing techniques of marine tardigrades?

I want to culture the tardigrades collected from intertidal area? In the initial experiment, I tried with diatom feed(Achnanthes) but failed.

30 April 2016 5,744 2 View

Vitro culture technique?

An isolated ovary is grown by which vitro culture?

06 April 2016 6,290 3 View

Vitro culture technique?

An isolated ovary is grown by which vitro culture?

04 April 2016 4,082 3 View

How can I avoid the granules like structure on the surface of the cells?

Hello Everyone,We are facing a problem in all cell lines running in our culture room. After two or three passages, the cells start showing the granules like structure on its surface. They...

04 April 2016 1,541 4 View

Can anyone suggest the culturing techniques of marine tardigrades?

I want to culture the tardigrades collected from intertidal area? In the initial experiment, I tried with diatom feed(Achnanthes) but failed.

04 April 2016 5,834 2 View

Which are the best anti-human CD44 neutralizing Abs for cell culture?

I'm looking for one Ab targeting CD44-HA interaction and one that doesn't. The aim is to use the Abs to disrupt CD44 adhesion/signaling in an in vitro tumor cell culture, eventually leading to...

04 April 2016 8,676 4 View

Primary human skin fibroblasts (HSF): what type of contamination could this be?

After several days in culture, rounded and refringent cells begin to appear. Over time, their number increases until they eventually die and detach from the flask's surface. Phase contrast images...

04 April 2016 5,722 2 View

Any suggestion on a positive control for CYP2D6?

Hello dear researchers, I already asked a similar question. I am looking for a cell line that can be used as a positive control for Western blot and for activty/inhibition assay for CYP2D6....

04 April 2016 2,553 1 View

I was wondering if people had recommendations on methods for using hTERT with Lenti SV 40Tt or HPV 16 E6/E7?

I am in the process of attempting to immortalize some primary cells. I used hTERT alone without success. Now I am usingLenti SV 40Tt and then Lenti HPV 16 E6/E7. With each of these treatments I am...

04 April 2016 1,254 2 View

Culturing mouse ES cells under feeder independent conditions- can anyone help?

Hi, I have purchased VGB6 mouse embryonic stem cells from Knockout Mouse Project (KOMP) Repository. These cells have been reported to be feeder-dependent. Does anyone have experience in culturing...

04 April 2016 7,832 9 View

Cell aggregation with HCT116?

Hi, I had a lot of problems with this cell line. When I count the number of cells, I can't do it, because the cells form a lot of cell aggregations and i can't separate them. The cells was...

03 March 2016 8,169 6 View

What is a good way to maintain Pythium irregulare?

I got good growth on Sabouraud agar with Pythium irregulare. Apparently P. irregulare (according to literature) shouldn't be freezed. So, after getting growth on agar plate (or slant) should I...

03 March 2016 6,536 0 View

How can I establish MEFs as a cell line?

As the title says, does it work for serial passage that MEFs transform and become immortal ?

03 March 2016 9,344 1 View

Expressing GFP in HEK and HeLa. no ECL results?

I have been working with GFP that I have exprrssed in HEK and HeLa cell lines. On transfer I can only see the marker on the nitrocellulose. I cannot even see the positive control on the...

03 March 2016 654 5 View

Can anyone please suggest to me a method for isolating cell membranes?

I am working with cell lines. As part of my work, I wish to isolate the membranes of cells. I wish to know if there is any simple method for cell membrane isolation which can be done without using...

03 March 2016 6,598 1 View

What can be the cause for contamination of cell culture?

Hello, I’m currently suffering a problem with my cells. The cells in question were recently transduced with lentivirus to obtain cells stably expressing a construct. After several weeks of...

03 March 2016 1,056 4 View

Anyone having BAC clone duplicator apparatus (equipment) usage expriences?

I am going to start a BAC clone multiplication. There is about 58600 clones kept at -80 degree C. in 384 well plate. I am searching for some 7 genes of my interest whereby I group clones and...

03 March 2016 551 0 View

Can anyone please suggest to me a method for isolating cell membranes?

I am working with cell lines. As part of my work, I wish to isolate the membranes of cells. I wish to know if there is any simple method for cell membrane isolation which can be done without using...

02 March 2016 9,584 1 View

Can laminin-coated culture vessels be reused for multiple cell passaging?

Do multiple times of cell dissociation inactivate the laminin activity? If the laminin-coated culture vessels can be reused several times, what is the limit to the number of times?

02 February 2016 421 3 View

Which tag is better to use for in vivo study in zebrafish; by Western blot or IP? Myc, FLAG, or HA? and how many copies should be used?

I want to do some in vivo study using zebrafish but I don't know which tag is better to use it for WB or IP and which one does not cause a background? Myc, Flag or HA?..Thanks in advance.

02 February 2016 9,910 2 View

Why are my cells over-confluent in the center of my dishes or wells?

I’ve seeded 6-well, 12-well, and 100mm dishes and continually find a heterogeneous distribution of my microvascular endothelial cells (primary cell line), regardless of seed density or technique...

02 February 2016 2,713 7 View

Is there a cell line to specifically study murine subcutaneous adipose tissue?

Hi, I am currently working on 3T3-L1 cell line to study subcutaneous adipose tissue (SAT). As my project is based on a specific effects on SAT (no effects on visceral adipose tissue) I am not...

01 January 2016 7,353 1 View

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