Isolation and transplantation of murine intestinal stem cells

This study has determined the location of the intestinal stem cells by labeling DNA with 5-bromo, 2^′deoxyuridine (BrdU). Based on the assumption that stem cells divide at a slower rate than their proliferating progeny, following a labeling period, cells are permitted to divide. With each cell division, DNA is replicated and the concentration of the pre-existing incorporated BrdU is halved. All rapidly dividing cells dilute the label at a greater rate than the slower dividing stem cells. These stem cells, highly concentrated with BrdU, are then identified. Each crypt was observed to contain one stem.

In order to bulk purify, a non-enzymatic manipulation of the gut was performed. This purification resulted in the release of viable epithelial cells in a sequential manner so as to reflect the gradient of cells from the villus tip to the crypt base.

Fourteen days post-transplant, the tissue was retrieved and the regeneration of intestinal morphology was assessed by the phenotypic characteristics of the newly transplanted cells. (Abstract shortened by UMI.)