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The effect of meconium exposure on the expression and differentiation of amniotic fluid mesenchymal stem cells
T J Jensen1, J E Shui2, C M Finck3
1Department of Pediatrics, UConn Health, Farmington, CT, USA.
Background:
The goal of this study was to determine if exposure to meconium would alter the phenotype of amniotic fluid mesenchymal stem cells (AF-MSCs) and the ability of these cells to be differentiated into distal airway type cells.
Methods:
Meconium was collected, lyophilized and resuspended in PBS at 3 different concentrations (high, medium, and low). AF-MSCs were cultured in the presence of this meconium suspension for 8 hours and then analyzed for changes in gene expression. Additionally, AF-MSCs exposed to meconium were differentiated for 14 days using modified small airway growth medium (mSAGM) and gene expression was determined. As a spontaneous differentiation control, meconium exposed AF-MSCs were cultured in amniotic fluid stem cell medium (AF medium).
Results:
After 8 hours of exposure in culture, AF-MSCs had increased expression of distal airway genes aquaporin 5 (AQP5) and surfactant protein c (SPC) when cultured in AF medium containing meconium. These gene expression levels were similar to that of AF-MSCs that were differentiated in mSAGM for 14 days. Furthermore, there was an up regulation of pluripotency genes NANOG and OCT4 in response to low meconium concentration for 8 hours. Following 14 days of culture in mSAGM, there was an upregulation of TTF1, SPC and AQP5 expression in the control, as well as in the low and medium meconium exposed groups indicating that these cells were still able to be differentiated. High meconium concentration did, however, appear to influence the level of distal airway gene expression after 14 days in mSAGM. After 14 days in AF medium, there was significant downregulation in pluripotency and mesenchymal markers as well as distal airway gene expression in all groups.
Conclusion:
The phenotype of AF-MSCs is modulated by meconium exposure; however, the cells were still able to differentiate into distal airway gene and protein expression. This result supports the hypothesis that progenitor cells exist in the amniotic fluid and the presence of meconium may affect their initial phenotype. However, these cells were still able to be differentiated to a distal lung phenotype.
Insights
Meconium exposure alters amniotic fluid mesenchymal stem cells (AF-MSCs) phenotype but does not prevent differentiation into distal airway cells. These findings suggest progenitor cells in amniotic fluid can adapt to meconium presence.
Area of Science:
- Stem cell biology
- Developmental biology
- Neonatal medicine
Background:
- Amniotic fluid mesenchymal stem cells (AF-MSCs) are progenitor cells with potential therapeutic applications.
- Meconium exposure in utero is a significant event in neonatal care.
- Understanding the impact of meconium on AF-MSCs is crucial for assessing fetal lung development.
Purpose of the Study:
- To investigate how meconium exposure affects the phenotype of AF-MSCs.
- To determine if meconium-exposed AF-MSCs retain their differentiation capacity into distal airway cells.
Main Methods:
- AF-MSCs were exposed to varying concentrations of meconium (high, medium, low) for 8 hours.
- Gene expression analysis was performed after initial meconium exposure.
- Meconium-exposed AF-MSCs underwent a 14-day differentiation protocol towards distal airway cells using modified small airway growth medium (mSAGM).
Main Results:
- Short-term meconium exposure (8 hours) increased distal airway gene expression (AQP5, SPC) and pluripotency genes (NANOG, OCT4) in AF-MSCs.
- AF-MSCs exposed to low and medium meconium concentrations successfully differentiated into distal airway cells, showing upregulation of TTF1, SPC, and AQP5 after 14 days.
- High meconium concentration impacted distal airway gene expression levels after 14 days, while prolonged culture in AF medium without mSAGM led to downregulation of key markers.
Conclusions:
- Meconium exposure modulates the phenotype of AF-MSCs, including initial changes in pluripotency and distal airway gene expression.
- Despite phenotypic modulation, AF-MSCs retain the ability to differentiate into cells expressing distal airway markers.
- These findings support the presence of adaptable progenitor cells in amniotic fluid that can respond to environmental cues like meconium.
