I used live alkaline phosphatase to prove that the adult stem cells were reprogrammed to more pluripotent cells. This was accompanied by a significantly lower expression of 3-germ layers differentiation markers. However, one comment was that alkaline phosphatase is also a marker for osteogenic differentiation. Is this comment valid?
Alkaline phosphatase is a perfectly valid pluripotent stem cell marker: Article Alkaline Phosphatase in Stem Cells
. Various alk phos isozymes are also found in a number of differentiated cell types, including osteogenic cells.
Altough Alk Phosp (AP) is a good marker for pluripotent cells (enzymatic activity), the comment is valid, and AP is also active in blood vessel.
Therefore, you could use other markers, together with AP, to unambigously demonstrate that cells are reprogrammed to pluripotent state: Immunostaining/immunofluorescence (IF) for NANOG (or VENTX) proteins (localization into the nucleus). Such a IF should be conveincing in order to claim "cells are reprgrammed to pluripotent STATE".
fIf you want to demonstrate that reprogrammed cells are FUNCTIONALLY pluripotent, you should differentiate them into 3 germ layers (ectoderm, mesoderm, endoderm, if you are using human cells) or to make a chimera (by injecting them into the blastocyst, if you are using mouse or other non-primate cells).
Alkaline phosphatase is an important component in hard tissue formation, highly expressed in mineralized tissue cells. AP expression increases when the stem cells are differentiated towards bone. Hence the comment about AP is valid.
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