Recovery of melanocytes - Seperation of epidermis from dermis and recovery of melanocytes (Jul/08/2009 )

I am a surgeon.
I see patients whose some areas of skin have white patches (vitiligo). This is very embarrassing condition and very striking in colored people.

Some of these patients can be treated by transferring their own melanocytes from other areas of the body.

A skin graft is taken and melanocytes are harvested from it and transferred to white patches after roughening it using diamond dust coated burrs.

I can take a skin graft and can rough up the white patch but I need to know the process of getting melanocytes from patient's own skin.

Now I request help in the following:

1.Seperation of epidermis from dermis and
2.Getting melanocytes from epidermis

I have seen a few surgeons practicing this art at conferences but are keeping this art of extraction of melanocytes very close to their chests.

they are using Trypsin/EDTA GMEM and soyabeen oil. They are using incubators and agitators. These are the only clues I have. These sound very gobbledygook to me but am willing to learn.

Can anyone help?
I am from India and there are enough sufferers who need help.

Earlier I used to transfer entire skin to roughened up white patch. But Melanocyte transfer is giving very good pleasing results in majority of patients.

I request moderators to move to appropriate forum

Hello apsara, and welcome to the BioForum!

I've moved your post here, as I think the Tissue and Cell Culture aficionados might have the best insight to your question. You're right that it doesn't fit perfectly anywhere...

I've also changed the title to be a bit more indicative of the information you're seeking. Hopefully, someone here will have some insight for you (I'm not a tissue guy; I work with bacteria).

sounds to me like DIY skin ( aka epidermal equivalets)

making this with a high enough level of melanocytes would be pretty cutting edge tho - maybe it is just a case of trypsinising the tissue to remove the dermis and pasting it on

not sure why you would want to separate off the melanocyes tho unless you were injecting them - a graft would need keritinocyes as well wouldnt it?

dom

apsara on Jul 8 2009, 03:38 PM said:

Hi Apsara

Dispase II helps in separation of epidermis from dermis. Alternatively you can use trypsin, which is faster. It is a standard procedure and you will find the exact conditions in a number of papers. I used to do it in my previous lab but don't remember the concentrations. For getting melanocytes, tease the epidermis nicely and plate the cells in M2 media, which is a media meant for culturing melanocytes alone. Alternatively, cascade biologicals have a media that is cheaper.

Many papers talk about MCDB153 with cholera toxin but I am not sure if the cells grown in this media can be used for transplantation due to cholera toxin or TPA.
Also look at the attached paper.

Hope this helps.

Best,
TC