Woww tq for this step by step video.. Very good effort 👍👏👏

Thank you for hand on practice video.

@MedicinalChemistryCenter

3 жыл бұрын

Thank you

This was a great, informative step-by-step video. Thank you for uploading. Quick question - is the interior of the culture flask coated with something to keep the cells adhered? It seems they don't come off until treatment with trypsin?

@MedicinalChemistryCenter

3 жыл бұрын

Hi Donne! Thanks for the comment. The flasks are not coated with the carrier. online-shop.eppendorf.ru/RU-ru/Kulturalnyj-plastik-110320/Rashodnye-materialy-dlja-raboty-s-kulturami-kletok-110321/Eppendorf-Cell-Culture-Flasks-PF-68138.html

@donneestipona1720

3 жыл бұрын

@@MedicinalChemistryCenter thank you for clarifying!

As a second question, if you are trying to dissociate spheroids (of mouse astrocytes) embedded in collagen (2.4mg/ml) matrix, what working soln concentration would you recommend for collagenase? Also, would it be beneficial to use a cocktail of collagenase, trypsin, and acutase in this case?

@MedicinalChemistryCenter

Жыл бұрын

Unfortunately, we have no experience with astrocytes, but your cocktail of enzymes seems to me a tough option for the dissociation of spheroids. I think a mixture of collagenase and acutase will be enough. Good luck, MCC

Hey, thank you for the detailed video. I have one question. When you add collagenase you mention it as 10mL of 60U/mL concentration But, when you are adding it to the small tissue pieces (present in the 50mL tube) you add two things using the pipette. Also, I understand you have prepared 60U/mL collagenase from the stock. Do you add it at that concentration directly to the cells or do you again dilute it while adding? Thank you!

@MedicinalChemistryCenter

Жыл бұрын

Dear Abhinaba Banerjee, We use ready-made collagenase solution in the laboratory (we use DMEM/F12 basal medium to dissolve the dry enzyme). The second reagent is calcium chloride, which is important for the work of collagenase. Good luck, MCC

@abhinababanerjee683

Жыл бұрын

@@MedicinalChemistryCenter Thank you for your reply! Also, from the stock to the working concentration do we dilute it in cell culture media? For the stock we use HBSS.

I noticed your bare wrists/arms and moving hands over the open samples - is there no risk of contamination in this kind of work? I'm a total beginner trying to understand the rules :)

@MedicinalChemistryCenter

3 жыл бұрын

There is no risk of contamination. Most importantly, wash your hands well up to the elbows before starting work.

what is the function of cacl2 and collagenase

@MedicinalChemistryCenter

2 жыл бұрын

Thank you for your question. Collagenase is used to de-internationalize tissue and isolate tumor cells. Calcium ions are essential for collagenase to function.