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406: Amniotic fluid derived cells: genome-wide expression in early and late cultures
www.AJOG.org
Doppler Assess, Fetus, Prematurity, U/S, Med-Surg-Diseases
406 Amniotic fluid derived cells: genome-wide expression in early and late cultures Ashwin Jadhav1, Jiri Zavadil3, Ross Basch3, Bruce Young2 1
UMDNJ-RWJMS, Obstetrics and Gynecology, New Brunswick, NJ, 2New York University Medical Center, Obstetrics and Gynecology, New York, NY, 3 New York University Medical Center, Department of Pathology, New York, NY
OBJECTIVE: Evaluates change in the genome wide expression profile of Amniotic Fluid Stem Cells during in-vitro culture STUDY DESIGN: Amniotic Fluid Stem Cells (AFSC) expressing CD117 (c-kit) surface markers were sorted using magnetic beads technique, mRNA extracted and converted to cDNA and in vitro transcribed to biotin-labeled cDNA using the standard Affymetrix 3’IVT Express Kit protocol. The labeled cDNA hybridization to the Affymetrix U133 2.0 arrays, array fluidics processing and scanning were performed under standardized conditions recommended by Affymetrix. Analysis of the microarray expression profile was performed using genespring multiomics software. RESULTS: We found that only 3.08% of gene probes were differentially expressed from early to late passage of AFSC culture. The differentially expressed genes were related to biological processes or cellular function - including transcription factors, protein kinases, and cytokines/ growth factors. Other gene-sets of interest were oncogenes and tumor suppressor genes, which were a very small number of genes. We further analyzed the gene sets of interest using NIH DAVID and GSEA bioinformatics databases for gene annotations analysis. Applying false discovery rate correction, there was no significant difference in the genome-wide expression profiling between early and late passage. CONCLUSION: Amniotic fluid derived stem cells (AFSCs) maintain their genome-wide expression profile during in-vitro culture.
Poster Session III
model to determine the effects of preeclampsia on fetal heart and lung development. STUDY DESIGN: Our experimental design included three groups: conventional-RUPP, with banding of the abdominal aorta and the ovarian arteries (cRUPP; n⫽3); modified-RUPP (mRUPP; n⫽4), with banding of the ovarian arteries and the uterine arteries; control (Ctr; n⫽4), with sham operation. Maternal hypertension and proteinuria were used to confirm preeclampsia. Surgery was performed on day 14 of gestation and on day 21 fetuses (n⫽91) were collected and assessed for reabsorption rates, fetal body, lung and heart weights. Lungs from each group were stained with hematoxylin-eosin and 10 fields from 2-3 sections per specimen were used for morphometric evaluation using the ImageJ 1.46 (NIH) software. RESULTS: Lung weights in the cRUPP (106⫾6 mg); and mRUPP (104⫾4 mg) groups were significantly lower (p⬍0.001) than controls (137⫾4 mg). The cRUPP group had also lower (p⬍0.04) body (4.0⫾1.0 g vs 4.3⫾0.6 g) and heart weights (27⫾1 mg vs 30⫾1 mg) compared to controls. Both cRUPP (56.2⫾15.3%, p⬍0.04) and mRUPP (81.4⫾2.2%; p⬍0.0002) had higher fetal reabsorption rates than controls (3.4⫾2.4%). Morphometric analysis also showed increased saccular area in the cRUPP (60.3⫾1.6%, p⬍0.01) and mRUPP (62.1⫾2.2%, p⬍0.005) groups compared to controls (54.7⫾.1.1%), and decreased saccular density expressed as mean counts/field (cRUPP only, 30⫾.2 vs 36⫾.2; p⬍0.05), confirming reduced lung growth associated with reduced lung complexity and maturation in the cRUPP group. CONCLUSION: Our data clearly indicate that preeclampsia has severe effects on fetal mortality and growth and that these effects are associated with fetal heart and lung growth restriction.
408 Pregnancies complicated with fetal growth restriction (FGR) is associated with a high rate of subsequent development of preeclampsia?
AFSC: differential gene expression analysis from early to late passage
Beth Dektas1, Baha Sibai2, Mounira Habli1 1 Good Samaritan Hospital, Obstetrics, Cincinnati, OH, 2UT houston, Obstetrics, Houston, TX
407 Fetal heart and lung growth restriction associated with preeclampsia Benedetta Pallante1, Mauro Schenone2, Federico Fernandez Nievas1, Giancarlo Mari2, Thomas Chin1 1
University of Tennessee Health Science Center, Pediatrics, Memphis, TN, University of Tennessee Health Science Center, Obstetrics and Gynecology, Memphis, TN 2
OBJECTIVE: Babies born from mothers with preeclampsia have increased risks of cardiovascular disease and chronic lung disease. The ethiopathogenesis could involve fetal heart and lung growth restriction resulting from reduced nutrients and/or oxygen supply to the fetus. We used a Reduced Uterine Pulse Pressure (RUPP) rat
OBJECTIVE: There are limited data regarding the rate of subsequent development of preeclampsia in pregnancies complicated by isolated FGR. Our objectives are to assess the rate of subsequent preeclampsia and the latency to development of preeclampsia in pregnancies complicated by isolated FGR. STUDY DESIGN: A retrospective cohort study of all singleton pregnancies (n⫽181) who were initially diagnosed with FGR at ⬍37 weeks (wks) with no clinical preeclampsia were studied. FGR was defined as ⬍10 % with negative work up for infection and karyotype. Exclusion criteria included preexisting hypertensive disorders, history of preeclampsia. Preeclampsia was defined per ACOG. Maternal and neonatal complications were evaluated. Data were stratified according to presence (cases) or absence of subsequent preeclampsia (control) and to gestational age(GA) at diagnosis of FGR (ⱕ32 vs. ⬎32 wks). RESULTS: 28(15%) developed subsequent preeclampsia at an average latency period of 1.38 ⫾2.3 wks (range 0.5-9.3). As compared to control (table), cases were diagnosed and delivered at an earlier GA with higher rates of both NICU admission and composite adverse neonatal outcome. As compared to FGR diagnosed at ⬎32 weeks (n⫽156), FGR diagnosed at ⱕ 32 weeks (n⫽25)had a significantly higher rate of subsequent preeclampsia (36%(9/25) vs. 12%(19/156), p⬍0.05), earlier GA at delivery (28.5⫾2.1 vs. 36.6⫾1.1 wks, p⬍0.001) but no difference in latency for development of preeclampsia (0.6⫾2.1 vs. 1.4⫾2.5 wks, p⬎0.05). CONCLUSION: Pregnancies complicated by isolated FGR are at increased risk of subsequent preeclampsia at an average of 1-2 weeks. The rate of subsequent progression of preeclampsia is dependent on gestational age at diagnosis. In addition, pregnancies complicated by subsequent preeclampsia are associated with increased adverse maternal and perinatal outcomes. We recommend that such pregnancies
Supplement to JANUARY 2013 American Journal of Obstetrics & Gynecology
S179
Doppler Assess, Fetus, Prematurity, U/S, Med-Surg-Diseases
406 Amniotic fluid derived cells: genome-wide expression in early and late cultures Ashwin Jadhav1, Jiri Zavadil3, Ross Basch3, Bruce Young2 1
UMDNJ-RWJMS, Obstetrics and Gynecology, New Brunswick, NJ, 2New York University Medical Center, Obstetrics and Gynecology, New York, NY, 3 New York University Medical Center, Department of Pathology, New York, NY
OBJECTIVE: Evaluates change in the genome wide expression profile of Amniotic Fluid Stem Cells during in-vitro culture STUDY DESIGN: Amniotic Fluid Stem Cells (AFSC) expressing CD117 (c-kit) surface markers were sorted using magnetic beads technique, mRNA extracted and converted to cDNA and in vitro transcribed to biotin-labeled cDNA using the standard Affymetrix 3’IVT Express Kit protocol. The labeled cDNA hybridization to the Affymetrix U133 2.0 arrays, array fluidics processing and scanning were performed under standardized conditions recommended by Affymetrix. Analysis of the microarray expression profile was performed using genespring multiomics software. RESULTS: We found that only 3.08% of gene probes were differentially expressed from early to late passage of AFSC culture. The differentially expressed genes were related to biological processes or cellular function - including transcription factors, protein kinases, and cytokines/ growth factors. Other gene-sets of interest were oncogenes and tumor suppressor genes, which were a very small number of genes. We further analyzed the gene sets of interest using NIH DAVID and GSEA bioinformatics databases for gene annotations analysis. Applying false discovery rate correction, there was no significant difference in the genome-wide expression profiling between early and late passage. CONCLUSION: Amniotic fluid derived stem cells (AFSCs) maintain their genome-wide expression profile during in-vitro culture.
Poster Session III
model to determine the effects of preeclampsia on fetal heart and lung development. STUDY DESIGN: Our experimental design included three groups: conventional-RUPP, with banding of the abdominal aorta and the ovarian arteries (cRUPP; n⫽3); modified-RUPP (mRUPP; n⫽4), with banding of the ovarian arteries and the uterine arteries; control (Ctr; n⫽4), with sham operation. Maternal hypertension and proteinuria were used to confirm preeclampsia. Surgery was performed on day 14 of gestation and on day 21 fetuses (n⫽91) were collected and assessed for reabsorption rates, fetal body, lung and heart weights. Lungs from each group were stained with hematoxylin-eosin and 10 fields from 2-3 sections per specimen were used for morphometric evaluation using the ImageJ 1.46 (NIH) software. RESULTS: Lung weights in the cRUPP (106⫾6 mg); and mRUPP (104⫾4 mg) groups were significantly lower (p⬍0.001) than controls (137⫾4 mg). The cRUPP group had also lower (p⬍0.04) body (4.0⫾1.0 g vs 4.3⫾0.6 g) and heart weights (27⫾1 mg vs 30⫾1 mg) compared to controls. Both cRUPP (56.2⫾15.3%, p⬍0.04) and mRUPP (81.4⫾2.2%; p⬍0.0002) had higher fetal reabsorption rates than controls (3.4⫾2.4%). Morphometric analysis also showed increased saccular area in the cRUPP (60.3⫾1.6%, p⬍0.01) and mRUPP (62.1⫾2.2%, p⬍0.005) groups compared to controls (54.7⫾.1.1%), and decreased saccular density expressed as mean counts/field (cRUPP only, 30⫾.2 vs 36⫾.2; p⬍0.05), confirming reduced lung growth associated with reduced lung complexity and maturation in the cRUPP group. CONCLUSION: Our data clearly indicate that preeclampsia has severe effects on fetal mortality and growth and that these effects are associated with fetal heart and lung growth restriction.
408 Pregnancies complicated with fetal growth restriction (FGR) is associated with a high rate of subsequent development of preeclampsia?
AFSC: differential gene expression analysis from early to late passage
Beth Dektas1, Baha Sibai2, Mounira Habli1 1 Good Samaritan Hospital, Obstetrics, Cincinnati, OH, 2UT houston, Obstetrics, Houston, TX
407 Fetal heart and lung growth restriction associated with preeclampsia Benedetta Pallante1, Mauro Schenone2, Federico Fernandez Nievas1, Giancarlo Mari2, Thomas Chin1 1
University of Tennessee Health Science Center, Pediatrics, Memphis, TN, University of Tennessee Health Science Center, Obstetrics and Gynecology, Memphis, TN 2
OBJECTIVE: Babies born from mothers with preeclampsia have increased risks of cardiovascular disease and chronic lung disease. The ethiopathogenesis could involve fetal heart and lung growth restriction resulting from reduced nutrients and/or oxygen supply to the fetus. We used a Reduced Uterine Pulse Pressure (RUPP) rat
OBJECTIVE: There are limited data regarding the rate of subsequent development of preeclampsia in pregnancies complicated by isolated FGR. Our objectives are to assess the rate of subsequent preeclampsia and the latency to development of preeclampsia in pregnancies complicated by isolated FGR. STUDY DESIGN: A retrospective cohort study of all singleton pregnancies (n⫽181) who were initially diagnosed with FGR at ⬍37 weeks (wks) with no clinical preeclampsia were studied. FGR was defined as ⬍10 % with negative work up for infection and karyotype. Exclusion criteria included preexisting hypertensive disorders, history of preeclampsia. Preeclampsia was defined per ACOG. Maternal and neonatal complications were evaluated. Data were stratified according to presence (cases) or absence of subsequent preeclampsia (control) and to gestational age(GA) at diagnosis of FGR (ⱕ32 vs. ⬎32 wks). RESULTS: 28(15%) developed subsequent preeclampsia at an average latency period of 1.38 ⫾2.3 wks (range 0.5-9.3). As compared to control (table), cases were diagnosed and delivered at an earlier GA with higher rates of both NICU admission and composite adverse neonatal outcome. As compared to FGR diagnosed at ⬎32 weeks (n⫽156), FGR diagnosed at ⱕ 32 weeks (n⫽25)had a significantly higher rate of subsequent preeclampsia (36%(9/25) vs. 12%(19/156), p⬍0.05), earlier GA at delivery (28.5⫾2.1 vs. 36.6⫾1.1 wks, p⬍0.001) but no difference in latency for development of preeclampsia (0.6⫾2.1 vs. 1.4⫾2.5 wks, p⬎0.05). CONCLUSION: Pregnancies complicated by isolated FGR are at increased risk of subsequent preeclampsia at an average of 1-2 weeks. The rate of subsequent progression of preeclampsia is dependent on gestational age at diagnosis. In addition, pregnancies complicated by subsequent preeclampsia are associated with increased adverse maternal and perinatal outcomes. We recommend that such pregnancies
Supplement to JANUARY 2013 American Journal of Obstetrics & Gynecology
S179