Primary Culture of Chicken Embryos - Step 4: Tissue Disaggregation

[@claudyomande2686]

what a beautiful demonstration (and honest with all the mess haha!) Thanks for making this video Professor Drew, looking forward to many more cell culturing vids!

[@ProfessorDrewCollop]

Claudyo. Thanks for your kind words. Working in Biotech can sometime be very "biological". I hope my video educated and entertained.

[@rajputsankar7520]

Thank you so much sir for making such efforts. All of your 4 videos were really helpful and of incredible knowledge. With love from India.

[@ProfessorDrewCollop]

Happy to hear you found them useful.

[@funny11744]

@@ProfessorDrewCollop how many days it has the embryo ?

[@ShivTea]

What do you mean by high quality chunks when it come to deciding on the samples you choose for the heart?

[@ProfessorDrewCollop]

When cutting an organ, such as the heart, for explant, you want to have a plan in place before cutting. Often, students can just start cutting up the organ creating a slimy goo, instead of a solid chunk of organ. For explant technique, I like to have defined edges to the chunks, as opposed to a liquid mash at the edge. With a defined edge, cells tend to migrate out from the chunk when they exit mitosis. If the cells do not migrate out and adhere to the dish, you will not be successful. Also, with explants, it is better to have a few really good chunks, than a dish covered in mashed up goo.

[@flpmuneeb-ws3hd]

Really good video and you covered all types of disaggregations in it. One this is , try to show the complete result Thank you.❤

[@ProfessorDrewCollop]

Are you asking me to show you an image of a successful cell line?

[@simranbhangu5499]

Thanks for the video. Its very informative

[@funny11744]

What method would you use for bone marrow isolation from the femur of that small chicken embrio ? The femur is too small to use a needle to extract the bone marrow. What method are used ? Thanks

[@ProfessorDrewCollop]

I have no experience attempting to extract bone marrow from a chicken embryo. Sorry, I cannot help you on this one.

[@joserosagomes7008]

Dear Professor. Have you a full protocol ( pdf file) about all these methods of cell primary culture? Please.

[@ProfessorDrewCollop]

Hi Jose. There is a lab manual that contains these protocol; however, it is published by my College. I do not believe I can just send you a PDF version of it. I am very clear in the videos about the protocol and would be happy to answer any questions you might have, if you try to create your own protocol while watching the videos. Sorry I cannot be more help.

[@abayomiruthtitilayo4922]

Can you do a video of growing normal edible egg from scratch in the lab. Like ivf you take the egg from the hen, but you growing the egg in the lab without fertilizing it.

[@ProfessorDrewCollop]

If the egg is not fertilized there will be nothing to grow. That is what an edible egg is, an unfertilized one.

[@abayomiruthtitilayo4922]

@@ProfessorDrewCollop yes that's what I want. Edible eggs. Can you teach one how to grow it. 😇😇😇

[@ProfessorDrewCollop]

This would be a question to ask an egg farmer.

[@abayomiruthtitilayo4922]

@@ProfessorDrewCollop I know, but I thought scientist now have a way to grow edible eggs that is not fertilized. Guess it's my imagination, I was just so interested in most of your videos. That's why asked if it's possible to have a edible egg in a lab without a hen. But since it's not possible, it's fine. Thank for responding back, I really appreciate.

[@ProfessorDrewCollop]

There is a process to grow edible meat in the lab now, without killing animals. Search for "Clean Meat" or "Cultured Meat" to get more inform. A process to replace the creation of an edible egg is not cost effective. Hens are cheap and not worth the effort.

[@alicehollingsworth8567]

where did you get the glucose from you milk vids (for positive control) all I can find is glucose powder but yours seems to be liquid?

[@ProfessorDrewCollop]

Thanks for the question Alice. We used reagent grade Glucose from Sigma and dissolved it in dH2O to a final concentration of 1 % (w/v).

[@HannahMckeown-nh1ft]

Hi there, wondering how long you leave the heart tissue in the fridge with the trypsin? And what are the steps that follow for this method? Having no luck with the transplant method and want to try this out.

[@ProfessorDrewCollop]

Hi Hannah. Thanks for the question. In the past we had our students only leave them in cold trypsin overnight. They needed to come back the following day to complete Day 2. As Lab is scheduled with 3 day break in between labs, we tried them for 3 days in the fridge. Still worked to make a monolayer of cells after incubation at 37'C for 30 min. No problems. I believe the trick is cut the tissue up into very small chunks to increase the surface area. Let me know how it goes.

[@funny11744]

Very interesting.

[@shirishzambre5751]

Very helpful