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

The subject line uses the term "therapeutic reinjection." Does this mean you're attempting to put the ASCs back into your body? If so, will you be testing the culture prior to injection for endotoxins? Have you planned on measuring the secretome of the cells, since ASCs have a tendency to produce proinflammatory cytokines (ILs 6, 7, 8 & 11, and TNF-alpha)? What is your intended biological endpoint for the therapy?

On Friday, December 18, 2020 at 11:10:38 AM UTC-5 fgarc...@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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