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Author: Subject: Is it possible to make a mammalian cell culture lab at home without breaking the bank? + 3 other questions
Nate
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[*] posted on 4-7-2018 at 21:43
Is it possible to make a mammalian cell culture lab at home without breaking the bank? + 3 other questions



2. I found on an article saying it's possible to use bicarbonate in place of a co2 incubator, is that true? (To be fair I did do a rough read)
https://www.researchgate.net/post/Can_mesenchymal_stem_cells...

3. Is it possible to make plasmids at home? If so, please provide insight on how this is done.

4. Any "cool" bacteria/fungus I could grow at home?
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Tsjerk
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[*] posted on 5-7-2018 at 04:34


Forget about the mammalian cell lines, you need both a proper lab and properly educated personal.

Micro-biology can be fun though, start of with isolation of E. coli with lactose, and distinguishing eukaryote and prokaryote using a microscope.

Plasmids could in theory be made in a home setting, but as far as I know there is no one who ever succeeded.
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Metacelsus
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[*] posted on 5-7-2018 at 17:41


As Tsjerk stated, mammalian cell culture in a home setting will be quite difficult. The growth medium is generally quite expensive. There are certain formulations for CO2-independent culture, but these tend to not work quite as well. Furthermore, the cells must be kept absolutely sterile, since any contamination will quickly and completely ruin your experiment.

I have experience with culturing (mouse) stem cells, and I wouldn't consider trying it at home. It simply wouldn't be worth the huge cost and effort to set up.

I have cultured yeast and bacteria at home. This is much easier. It wouldn't be too difficult to prepare a recombinant plasmid at home. The necessary enzymes and reagents would probably cost less than $100. Here's an example protocol for plasmid isolation: https://bitesizebio.com/13519/how-to-do-a-kit-free-midiprep/

For "cool" bacteria I would suggest luminescent Vibrio fischeri which can be isolated from raw seafood.







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Harristotle
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[*] posted on 5-7-2018 at 19:17


Hi. I used bicarbonate/citrate (ENO) in dishes to culture fish cells at low temperature, a long time ago. Yes it can be done, and works well too. I can't find my protocol, because I chucked that lab book out 30 years ago. I had a CO2 concentration measurer and made a standard curve- perhaps you could just do it volumetrically by measuring how much precipitate you get when you take 100ml of the air/CO2 mix and react with Ca(OH)2, or some other insoluble.

As I recall, a lot of the original cell culture work was done on white blood cells, using the "hanging drop method". This involved exposing the cells to a mitogen (ie bits of bacterial cell wall?), and culturing in a drop of serum. Serum being made by collecting whole blood and allowing it to clot in a tube. When the clot has formed and contracted, the clear yellow fluid above is the serum. It is an old technique, but worked. I am sure that for karyotyping using a bit of colchicine, it would do fine! Sterility is an issue, but a drop of serum in an oil droplet used to be used (if I remember correctly).

A bit fiddly for home, but doable. An absolute no-no for schools due to all the human fluids and disease protocols.

Cheers,
H.
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Nate
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[*] posted on 6-7-2018 at 09:58


Hi, thank you for all your responses! Tsjerk, may I ask how the isolation of e.coli with lactose is performe

edit: Harristotle, may you please provide more info on those old techniques please? Thank you in advanced!!

edit: Metacelsus, could you list the reagents and enzymes needed in that procedure? I read over it but some I'm not familiar with.

[Edited on 6-7-2018 by Nate]

[Edited on 6-7-2018 by Nate]
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