How many passages can we keep neonatal cardiomyocytes?

More Isehaq Al-Huseini's questions See All

Can we distinguish the type of free fluid inside the pelvic and abdominal cavities whether blood or serous fluid using the ultrasound?

during ultrasound examination of acute abdomen, sometimes we see free fluid , can we know the type of fluid ,whether blood or not , using ultrasound only? if yes , what are the sonographic...

01 March 2021 7,476 4 View

The reason that the culture of white fungi Contamination with green mold(Trichoderma), the culture autoclave at121 for 15 min. What is it?

Is the period to autoclave not enough? The inoculation in a hood and flame and UV and alcohol.

27 February 2021 9,356 3 View

Is there a fast method matching citations to bibliography (manually typed references)?

I need to be able to match in-text citations to a reference list and back again in very large documents (100+ pages) WITHOUT the use of referencing software like Endnote. my method is to type the...

27 February 2021 9,848 1 View

Can I put a figure that was drawn by Bio-Render in publication paper?

I want to publish paper in journal but I have figures which have drawn with bio-renders, can I put them in the paper or which other software I can use to draw them?

27 February 2021 8,179 1 View

What is the most suitable medium of ascomycetes fungi ??

26 February 2021 4,413 1 View

Is it possible to set a breakpoint at a specific DYNAMIC instruction using the GNU Debugger (GDB)?

Using GDB, it is straightforward to debug and monitor a target program by setting a break-point at a specific instruction, since the instruction addresses are known. However, we have two...

24 February 2021 1,598 2 View

Is it possible that Cancer cells could survive in heparinized blood for 48 hours?

Hi everyone, I am just wondering if we incubated 200 microliter of whole blood with 50 microliter of cancer cells(1bout 5000cells) do you think that cancer would survive for 48 hours?

23 February 2021 3,840 3 View

What is the most prevalent drug related problem in the developed countries?

Detecting the drug related problems is an important step in the pharmaceutical care plan

21 February 2021 5,421 3 View

What are the dimensions by which the independent variable (big data) can be measured, Knowing We using the questionnaire instrument?

We have a research project consisting of three variables where big data is the independent variable. Some previous studies showed that Volume, Variety, Velocity and Veracity Are the dimensions...

21 February 2021 8,133 2 View

Is nanotechnology part of materials science? Is it possible to obtain Pico technology and move from hypothesis to application?

Pico-scale This is three orders of magnitude smaller than a nanometer (and thus most nanotechnology) and two orders of magnitude smaller than most chemistry transformations and measurements. Pico...

18 February 2021 4,540 5 View

No title

02 March 2021 3,060 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Is there a method of determining feed digestibility without the need to re-pellet the feed?

I am aiming to determine the ADC values of a commercial fish feed, but I do not have a steam pelleter or extruder available to me. Ideally, the pellets could be coated with some inert marker....

28 February 2021 7,967 3 View

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

No title

28 February 2021 9,936 2 View

Bought HUVEC from Invitrgen.need to revive it.expected it is cryofreezed in DMSO.Do I need to remove the DMSO for plating the cell for the 1st time?

After I cryofreeze the cell do I need to remove the DMSO for culturing the cell for next passages .

28 February 2021 7,738 3 View

Saad Moghannem

good effort

please keep in consideration this is normal cells with the all characteristics of normal tissuebehavior. that means the number of passage not exceed 4 to 6 times. also preserve the cells with small number of passage as you can in vapour phase of liquid nitrogen. it should be with good viability for about one year if the preservation condition of liquid nitrogen tank is good.

also the number of preserved cells in cryovials should be more than 5 X 106 cell/ ml and resuscitated in small baby tissue culture flask

BR.,

Isehaq Al-Huseini

Thank you, Dr. Saad, for your informative answer

I am just wondering if this instruction is common for any primary cells or also can be applied for primary cardiomyocytes cell which looks more sensitive than other cells.

Have you applied this on primary neonatal cardiomyocytes cells

I mean keeping cells more than 4 passages and preserve them in liquid nitrogen for a long time.

Hélène Peyret

I frequently isolate beating cardiomyocytes from newborn rats, and unfortunately we are not able to make passages in culture. Indeed, the beating capacity of the cardiomyocytes is lost after the first passage. Have you noticed that in your culture?

Regards,

H PEYRET

I am working with different types of primary cultures from different organs liver, skin, kidney and spleen. Each types of culture has it's own behavior and characteristics even the same cell origin from different sources.

but I don't working with primary cardiomyocytes and I know this types of cells is very sensitive.

saad

Hi Hélène Peyret

Thank you for answering.

Yes, I noticed that in my cells, beating capacity is getting less at the time when they reach confluency. It might be better to harvest neonatal cardiomyocytes immediately after getting confluent.

Best regards

Isehaq

Yes I agree with you. Moreover, neonatal cardiomyocytes don't have a high proliferation capacity. After 2 or 3 passages, I think there are almost only fibroblasts in the culture, which are less sensitive and proliferate more rapidly. This should be verified by immunofluorescence (troponin I or T). In my lab, we use cardiomyocytes only 4 to 10 days after the first seeding and don't make any passage. Good luck !