Hypocapnia leads to enhanced expression of pluripotency and meso-endodermal differentiation genes in mouse embryonic stem cells.
The efficient utilization of embryonic stem cells for applications like cell based therapy, transplantation and drug discovery largely depends upon the culturing conditions of these cells. In this report, we have analyzed gene, protein expression and morphological changes of embryonic stem cells when subjected to lowered CO2 levels i.e. hypocapnia. We studied the quantitative expression of pluripotent genes, Oct3/4, Nanog and Sox2 and genes involved in the differentiation to the three lineages, under varying CO2 levels. Enhanced expression of these genes was seen at cultures maintained at 1.5% CO2 as compared to those maintained at 5% CO2. The cells exposed to hypocapnic conditions when subjected to immunocytochemical analysis stained positive for Oct-3/4, Nanog and Sox2 transcription factors. Flow cytometry and western blot further showed that the pluripotent proteins in the 1.5% CO2 maintained cultures have higher levels of expression as compared to the ES cells at 5% CO2. In addition, there was enhanced differentiation particularly towards the mesodermal and endodermal lineages at cultures maintained and differentiated at 1.5% CO2 at all the time periods analyzed i.e. day 10 (5+5d), 12 (5+7d) and day 15 (5+10d). These results, which we feel are the first of their kind, indicate that lowered CO2 levels seem to be preferred for the maintenance of pluripotency and the subsequent differentiation.
['Jyoti|Saras|S|', 'Tandon|Simran|S|']
D000469:Alkaline Phosphatase D000818:Animals D015415:Biomarkers D015153:Blotting, Western D002454:Cell Differentiation D019070:Cell Lineage D002478:Cells, Cultured D053595:Embryonic Stem Cells D004707:Endoderm D005434:Flow Cytometry D005455:Fluorescent Antibody Technique D018507:Gene Expression Regulation, Developmental D018398:Homeodomain Proteins D016857:Hypocapnia D008648:Mesoderm D051379:Mice D008856:Microscopy, Fluorescence D000071317:Nanog Homeobox Protein D050814:Octamer Transcription Factor-3 D039904:Pluripotent Stem Cells D012333:RNA, Messenger D060888:Real-Time Polymerase Chain Reaction D020133:Reverse Transcriptase Polymerase Chain Reaction D055748:SOXB1 Transcription Factors
Hypocapnia Pluripotency Differentiation Embryonic stem cells