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A gestational profile of placental exosomes in maternal plasma and their effects on endothelial cell migration.

Salomon C, Torres MJ, Kobayashi M, Scholz-Romero K, Sobrevia L, Dobierzewska A, Illanes SE, Mitchell MD, Rice GE - PLoS ONE (2014)

Bottom Line: The effect of exosomes isolated from FT, ST and TT on endothelial cell migration were established using a real-time, live-cell imaging system (Incucyte).During normal healthy pregnancy, the number of exosomes present in maternal plasma increased significantly with gestational age by more that two-fold (p<0.001).Pregnancy is associated with a dramatic increase in the number of exosomes present in plasma and maternal plasma exosomes are bioactive.

View Article: PubMed Central - PubMed

Affiliation: University of Queensland Centre for Clinical Research, Centre for Clinical Diagnostics, Royal Brisbane and Women's Hospital, Queensland, Australia.

ABSTRACT
Studies completed to date provide persuasive evidence that placental cell-derived exosomes play a significant role in intercellular communication pathways that potentially contribute to placentation and development of materno-fetal vascular circulation. The aim of this study was to establish the gestational-age release profile and bioactivity of placental cell-derived exosome in maternal plasma. Plasma samples (n = 20 per pregnant group) were obtained from non-pregnant and pregnant women in the first (FT, 6-12 weeks), second (ST, 22-24 weeks) and third (TT, 32-38 weeks) trimester. The number of exosomes and placental exosome contribution were determined by quantifying immunoreactive exosomal CD63 and placenta-specific marker (PLAP), respectively. The effect of exosomes isolated from FT, ST and TT on endothelial cell migration were established using a real-time, live-cell imaging system (Incucyte). Exosome plasma concentration was more than 50-fold greater in pregnant women than in non-pregnant women (p<0.001). During normal healthy pregnancy, the number of exosomes present in maternal plasma increased significantly with gestational age by more that two-fold (p<0.001). Exosomes isolated from FT, ST and TT increased endothelial cell migration by 1.9±0.1, 1.6±0.2 and 1.3±0.1-fold, respectively compared to the control. Pregnancy is associated with a dramatic increase in the number of exosomes present in plasma and maternal plasma exosomes are bioactive. While the role of placental cell-derived exosome in regulating maternal and/or fetal vascular responses remains to be elucidated, changes in exosome profile may be of clinical utility in the diagnosis of placental dysfunction.

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Maternal exosome effects on endothelial cells migration.HUVEC were grown to confluence in PCM and a wound was made using 96 well WoundMaker (see Methods). HUVEC Migration was measured in absence or presence of 100 ug/ml of exosomes from first, second and third trimester for 24 h. (A) Graphical representation from a showing the calculation of initial wound width (black) and graphical representation of cell migration (gray) at the midpoint of the experiment. (B) Time course of wound closure for HUVEC expressed as relative wound density (%). (C) Area under curves from data in B. Data represent an n = 12 well each point with 6 different cells culture (i.e. biological replicates) of HUVEC isolated from first trimester pregnancies (see methods). Values are mean ± SEM. In C, *p<0.05 versus all condition; †P<0.005 versus second and third trimester exosomes.
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pone-0098667-g006: Maternal exosome effects on endothelial cells migration.HUVEC were grown to confluence in PCM and a wound was made using 96 well WoundMaker (see Methods). HUVEC Migration was measured in absence or presence of 100 ug/ml of exosomes from first, second and third trimester for 24 h. (A) Graphical representation from a showing the calculation of initial wound width (black) and graphical representation of cell migration (gray) at the midpoint of the experiment. (B) Time course of wound closure for HUVEC expressed as relative wound density (%). (C) Area under curves from data in B. Data represent an n = 12 well each point with 6 different cells culture (i.e. biological replicates) of HUVEC isolated from first trimester pregnancies (see methods). Values are mean ± SEM. In C, *p<0.05 versus all condition; †P<0.005 versus second and third trimester exosomes.

Mentions: The effect of maternal circulation-derived exosomes isolated from first (FT-exo), second (ST-exo) and third (TT-exo) trimester of pregnancy on endothelial cell migration was determined using a real time cell imaging system (IncuCyte live-cell ESSEN BioScience Inc, Ann Arbor, Michigan, USA) and human umbilical vein endothelial cells (HUVEC). The effect of 100 µg exosomal protein/ml on HUVEC migration is presented in Figure 6 A, B and C. Representative photomicrographs of HUVEC wound closure with treatment (exo-FT, exo-ST and exo-TT) are presented in Figure 6A. The effect of exosomes on HUVEC migration is presented as relative wound density (percent) overtime (Figure 6B). Analysis of the area under the cell migration curves was used to assess the effect of gestational age on the bioactivity of maternal plasma exosomes. Maternal plasma exosomes isolated from FT, ST and TT increased HUVEC migration by: 2.7 fold; 2.3 fold; and 1.87 fold respectively when compared to cell migration without exosomes (-exo) (Figure 6C). In contrast, exosomes from non-pregnant women increases HUVEC migration ∼1.45 fold compared to cell migration without exosomes (-exo). The effect of exo-FT and exo-ST on HUVEC migration was significantly higher (p<0.05) versus the effect of exosomes from non-pregnant women. In addition, VEGF induced (∼2.9-fold) HUVEC migration to compare to control in absence of exosomes.


A gestational profile of placental exosomes in maternal plasma and their effects on endothelial cell migration.

Salomon C, Torres MJ, Kobayashi M, Scholz-Romero K, Sobrevia L, Dobierzewska A, Illanes SE, Mitchell MD, Rice GE - PLoS ONE (2014)

Maternal exosome effects on endothelial cells migration.HUVEC were grown to confluence in PCM and a wound was made using 96 well WoundMaker (see Methods). HUVEC Migration was measured in absence or presence of 100 ug/ml of exosomes from first, second and third trimester for 24 h. (A) Graphical representation from a showing the calculation of initial wound width (black) and graphical representation of cell migration (gray) at the midpoint of the experiment. (B) Time course of wound closure for HUVEC expressed as relative wound density (%). (C) Area under curves from data in B. Data represent an n = 12 well each point with 6 different cells culture (i.e. biological replicates) of HUVEC isolated from first trimester pregnancies (see methods). Values are mean ± SEM. In C, *p<0.05 versus all condition; †P<0.005 versus second and third trimester exosomes.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4048215&req=5

pone-0098667-g006: Maternal exosome effects on endothelial cells migration.HUVEC were grown to confluence in PCM and a wound was made using 96 well WoundMaker (see Methods). HUVEC Migration was measured in absence or presence of 100 ug/ml of exosomes from first, second and third trimester for 24 h. (A) Graphical representation from a showing the calculation of initial wound width (black) and graphical representation of cell migration (gray) at the midpoint of the experiment. (B) Time course of wound closure for HUVEC expressed as relative wound density (%). (C) Area under curves from data in B. Data represent an n = 12 well each point with 6 different cells culture (i.e. biological replicates) of HUVEC isolated from first trimester pregnancies (see methods). Values are mean ± SEM. In C, *p<0.05 versus all condition; †P<0.005 versus second and third trimester exosomes.
Mentions: The effect of maternal circulation-derived exosomes isolated from first (FT-exo), second (ST-exo) and third (TT-exo) trimester of pregnancy on endothelial cell migration was determined using a real time cell imaging system (IncuCyte live-cell ESSEN BioScience Inc, Ann Arbor, Michigan, USA) and human umbilical vein endothelial cells (HUVEC). The effect of 100 µg exosomal protein/ml on HUVEC migration is presented in Figure 6 A, B and C. Representative photomicrographs of HUVEC wound closure with treatment (exo-FT, exo-ST and exo-TT) are presented in Figure 6A. The effect of exosomes on HUVEC migration is presented as relative wound density (percent) overtime (Figure 6B). Analysis of the area under the cell migration curves was used to assess the effect of gestational age on the bioactivity of maternal plasma exosomes. Maternal plasma exosomes isolated from FT, ST and TT increased HUVEC migration by: 2.7 fold; 2.3 fold; and 1.87 fold respectively when compared to cell migration without exosomes (-exo) (Figure 6C). In contrast, exosomes from non-pregnant women increases HUVEC migration ∼1.45 fold compared to cell migration without exosomes (-exo). The effect of exo-FT and exo-ST on HUVEC migration was significantly higher (p<0.05) versus the effect of exosomes from non-pregnant women. In addition, VEGF induced (∼2.9-fold) HUVEC migration to compare to control in absence of exosomes.

Bottom Line: The effect of exosomes isolated from FT, ST and TT on endothelial cell migration were established using a real-time, live-cell imaging system (Incucyte).During normal healthy pregnancy, the number of exosomes present in maternal plasma increased significantly with gestational age by more that two-fold (p<0.001).Pregnancy is associated with a dramatic increase in the number of exosomes present in plasma and maternal plasma exosomes are bioactive.

View Article: PubMed Central - PubMed

Affiliation: University of Queensland Centre for Clinical Research, Centre for Clinical Diagnostics, Royal Brisbane and Women's Hospital, Queensland, Australia.

ABSTRACT
Studies completed to date provide persuasive evidence that placental cell-derived exosomes play a significant role in intercellular communication pathways that potentially contribute to placentation and development of materno-fetal vascular circulation. The aim of this study was to establish the gestational-age release profile and bioactivity of placental cell-derived exosome in maternal plasma. Plasma samples (n = 20 per pregnant group) were obtained from non-pregnant and pregnant women in the first (FT, 6-12 weeks), second (ST, 22-24 weeks) and third (TT, 32-38 weeks) trimester. The number of exosomes and placental exosome contribution were determined by quantifying immunoreactive exosomal CD63 and placenta-specific marker (PLAP), respectively. The effect of exosomes isolated from FT, ST and TT on endothelial cell migration were established using a real-time, live-cell imaging system (Incucyte). Exosome plasma concentration was more than 50-fold greater in pregnant women than in non-pregnant women (p<0.001). During normal healthy pregnancy, the number of exosomes present in maternal plasma increased significantly with gestational age by more that two-fold (p<0.001). Exosomes isolated from FT, ST and TT increased endothelial cell migration by 1.9±0.1, 1.6±0.2 and 1.3±0.1-fold, respectively compared to the control. Pregnancy is associated with a dramatic increase in the number of exosomes present in plasma and maternal plasma exosomes are bioactive. While the role of placental cell-derived exosome in regulating maternal and/or fetal vascular responses remains to be elucidated, changes in exosome profile may be of clinical utility in the diagnosis of placental dysfunction.

Show MeSH
Related in: MedlinePlus