Re: [DIYbio] An update on my effort to isolate viable adipose derived stem cells for therapeutic reinjection and a call for advice

Pretty interesting pictures in some of the articles. Amazing you can
take out adipose tissue, diggest it, spin it, and harvest a plethora
of cell types.

As you mentioned, it looks like only 5% of that final fraction is
adipose derived stem cells. I imagine it will be difficult to select
only the ADSC's from that fraction with low-budget equipment. A flow
cytometry machine would be able to do it. A very careful hand and a
tiny pulled glass syringe might under a microscope.

I've done 0 human cell line culture in my life but even the lab next
to us who has an ozonator, secure room, hoods, everything, often
complains of contamination of fungal spores etc. It's not easy!

You may just take the cell mixture found in the VSF and culture that
and see what cells tend to dominate in it. It may be the stem cells
do well in culture and out-grow other types. It may be they do not,
and further purification to a monoculture is needed. How you'd do
that, I have no idea but it's interesting.

On Fri, Dec 18, 2020 at 11:10 AM Frank Garcia <fgarcia0007@gmail.com> wrote:
>
>
> So I've scoured the internet for papers and guides to nonenzymatic procedures that can be done with simple inexpensive equipment and which yields an usable quantity of fat stem cells that are alive and well.
>
> I've read a few and must say I'm not entirely sure if I understand the required steps.
> Each of the following describes variations of the procedure. Almost all add some fancy element in one of the steps like a commercial kit, specialized syringe with filters built in , or custom made contraptions - get a load of this "Custom-made metallic disarmable pistons, concave cell-adhesive gaskets, closed cubic unit harnessing 3 different-sized sets of blade grids on each luer-lock port, rotating canal at the center of the cube to control the flow of the lipoaspirate, http://links.lww.com/PRSGO/B308).".
>
> https://www.nature.com/articles/s41598-017-10710-6#Tab3
> https://journals.lww.com/prsgo/fulltext/2020/02000/a_3_step_mechanical_digestion_method_to_harvest.27.aspx
> https://stemcells-aesthetica.com/isolation-process-of-stem-cells-from-fatty-tissue/
> http://www.irvinesci.com/protocol-for-mesenchymal-stem-cell-isolation
>
> I'd like to use these to put together a protocol that uses no expensive or custom made equipment. I'd like to know if the plating and incubation is necessary and is filtering necessary? Also, is the VSF the end point or can ADSCs be isolated from VSF? any other feedback welcome.
>
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