Exequibility of differential gene expression analysis by DDRT-PCR in murine bone marrow cells

Abstract

Model of study: Experimental study. Introduction: Recently, stem cell research has generated great
interest due to its applicability in regenerative medicine. Bone marrow is considered the most important
source of adult stem cells and the establishment of new methods towards gene expression analysis
regarding stem cells has become necessary. Thus Differential Display Reverse Transcription Polymerase Chain Reaction (DDRT-PCR) may be an accessible tool to investigate small differences in the gene
expression of different stem cells in distinct situations.
Aim: In the present study, we investigated the exequibility of DDRT-PCR to identify differences in global
gene expression of mice bone marrow cells under two conditions.
Methods: First, bone marrow cells were isolated fresh and a part was cultivated during one week without
medium replacement. Afterwards, both bone marrow cells (fresh and cultivated) were submitted to gene
expression analyses by DDRT-PCR.
Results: Initially, it was possible to observe in one week-cultured bone marrow cells, changes in morphology (oval cells to fibroblastic-like cells) and protein profile, which was seen through differences in
band distribution in SDS-Page gels. Finally through gene expression analysis, we detected three bands
(1300, 1000 and 225 bp) exclusively expressed in the fresh bone marrow group and two bands (400 and
300 bp) expressed specifically in the cultivated bone marrow cell group.
Conclusions: In summary, the DDRT-PCR method was proved efficient towards the identification of
small differences in gene expression of bone marrow cells in two defined conditions. Thus, we expect
that DDRT-PCR can be fast and efficiently designed to analyze differential gene expression in several
stem cell types under distinct conditions.