Effects of Pharmacological Agents on Human Amniotic Fluid-Derived Stem Cells in Culture

Abstract

Amniotic fluid contains multipotent cells and could be a source of stem cells for clinical use. Amniotic fluid cells (AFCs) are made up of a heterogeneous population of fetal cells that can be retrieved during pregnancy without ethical concerns as it is a standard clinical procedure. Studies of proliferation, multipotent marker expression, differentiation, and gene expression were performed after culturing with dexamethasone, valproic acid, and magnesium sulfate. There were correlations between known drug effects on the human fetus and changes seen in human AFCs in culture, as well as previously undescribed observations in neural and chondrogenic inducibility. Gene expression profiles confirmed these observations. AFC culture may provide a novel method to evaluate pharmacological agents before clinical use in pregnancy.

Methotrexate and Valproic Acid Affect Early Neurogenesis of Human Amniotic Fluid Stem Cells from Myelomeningocele

“…Concentrations of both pharmacological agents were setup considering previous studies [ 20 ] and dose-response experiments (Supporting Information Figures 1(a) and 1(b)). Increasing concentrations were tested for each pharmacological agent, and cell viability assay analyses were performed.…”

“…Subsequently, immature neurons expressing neural β III-tubulin and radial glial (RG) cells expressing BLBP appear for further neural tube organization [ 14 – 18 ]. FA stimulates the proliferation and differentiation of neural stem cells (NSCs), whereas MTX and VPA impair cell proliferation of embryonic NSCs and amniotic fluid-derived neural stem cells, respectively [ 19 , 20 ]. It is likely that undesirable effects of VPA and MTX on the developing neural tube are due to the influence of genes crucial for neural tube development, including Ki67, SOX2, and NESTIN [ 7 , 20 ].…”

“…FA stimulates the proliferation and differentiation of neural stem cells (NSCs), whereas MTX and VPA impair cell proliferation of embryonic NSCs and amniotic fluid-derived neural stem cells, respectively [ 19 , 20 ]. It is likely that undesirable effects of VPA and MTX on the developing neural tube are due to the influence of genes crucial for neural tube development, including Ki67, SOX2, and NESTIN [ 7 , 20 ]. Obviously, the assessment of undesirable effects of drugs in pregnancy is limited for ethical reasons, with limited options to enroll pregnant women in clinical trials [ 20 ].…”

“…It is likely that undesirable effects of VPA and MTX on the developing neural tube are due to the influence of genes crucial for neural tube development, including Ki67, SOX2, and NESTIN [ 7 , 20 ]. Obviously, the assessment of undesirable effects of drugs in pregnancy is limited for ethical reasons, with limited options to enroll pregnant women in clinical trials [ 20 ]. Thus, retrospective epidemiological studies or experimental animal models remain the main sources for studying drug effects.…”

“…AFSCs are a subset of multipotent fetal stem cells that retain the ability to differentiate various cell types [ 21 – 24 ]. Previous studies have demonstrated successful differentiation of AFSCs to neural-like cells [ 25 – 28 ], and pharmacological agents, including VPA, have been shown to affect their characteristics [ 20 ].…”