Stem Cell Niche - Murine Adult hematopoietic stem cell division Niche?? (Aug/09/2008 )

hi everybody, i m new to stem cell research... presently i m involved in sorting CD45 negative cells (stromal cells) from murine adult hematopoietic stem cells through MACS(miltenyi biotech) but not able to sort out the population. i also did FACS analysis of the negative sorted population. % of impurity was 80%. if anybody can suggest a proper protocol for sorting out the said population. I followed the standard protocol of MACS.
thanks in advance

---

The easiest thing to do would be to up the number of Ab's that you use to select out (negative selection) the cells you don't want. CD3, CD4, CD8, CD19, B220, Ter-119 can be used as a dump for excluding the cells you won't want to look at by Facs (sorting or analysis). Make them all PE labeled or some such color and stain your stem cells with c-kit and sca-1 or CD201 (which stains the same population as c-kit and sca-1 combined) and maybe CD34. There is a company called ebioscience that makes a cocktail or you can buy the Ab's individually. They are usually pretty cheap...most of the time much cheaper than Becton Dickinson.

www.ebioscience.com

---