Return to study ST001038 main page
MB Sample ID: SA069550
| Local Sample ID: | 12_MP_24 |
| Subject ID: | SU001077 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Human Inclusion Criteria: | Caesarean delivery, Non-labored, Full term |
| Human Exclusion Criteria: | Twins |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001077 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Human Inclusion Criteria: | Caesarean delivery, Non-labored, Full term |
| Human Exclusion Criteria: | Twins |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 12_MP_24 | SA069550 | FL011129 | 24 hr | Timepoint |
| 12_MP_24 | SA069550 | FL011129 | Maternal | Placenta_Surface |
| 12_MP_24 | SA069550 | FL011129 | Periphery | Placenta_Area |
Collection:
| Collection ID: | CO001071 |
| Collection Summary: | The placentas were collected immediately following cesarean delivery, and the tissue specimens were collected from the whole placenta at five time points postdelivery: (1) < 5 min (n = 6); (2) 15 min (n = 13); (3) 30 min (n = 13); (4) 1 h (n = 13); (5) 24 h (n = 13). The tissue specimens were immediately frozen in liquid nitrogen following collection. The whole placentas were kept at 4 °C between sampling time points. Tissue specimens were collected from two areas of the maternal (approximately 1 × 1 cm area; depth of 1 cm) and fetal surfaces (approximately 1 × 1 cm area; depth of 0.3 cm) of each placenta at each time point, resulting in 24 total specimens for the < 5 min time point and 52 total specimens for all other time points. Tissue specimens were immediately frozen in liquid nitrogen following collection and stored at −80 °C until metabolomic analysis |
| Sample Type: | Placenta |
| Collection Method: | Tissue specimens were collected from two areas of the maternal (approximately 1 × 1 cm area; depth of 1 cm) and fetal surfaces (approximately 1 × 1 cm area; depth of 0.3 cm) of each placenta at each time point by cutting with sterile scalpel and forceps on a sterile surface. The tissue specimens were immediately frozen in liquid nitrogen following collection. |
| Collection Frequency: | Tissue specimens were collected from the whole placenta at five time points postdelivery: (1) < 5 min (n = 6); (2) 15 min (n = 13); (3) 30 min (n = 13); (4) 1 h (n = 13); (5) 24 h (n = 13) |
| Storage Conditions: | -80℃ |
| Collection Vials: | Nunc 1.8mL Cryogenic tubes (ThermoFisher) |
| Storage Vials: | Nunc 1.8mL Cryogenic tubes (ThermoFisher) |
Treatment:
| Treatment ID: | TR001091 |
| Treatment Summary: | NA |
Sample Preparation:
| Sampleprep ID: | SP001084 |
| Sampleprep Summary: | The placental tissue samples (24.15–192.21 mg; mean 126.21 mg) were cut on dry ice and washed with ice-cold 0.85% sodium chloride to remove the excess blood before being placed in prechilled tubes. Five hundred μL of ice-cold 50/50 methanol/water was added to each sample before vortexing for 1 min. Homogenization and sonication methods were compared for metabolite extractions. Equivalent results were observed for both methods and, therefore, sonication was used because of its ability for high-throughput sample preparation. The samples were sonicated for 20 min to extract the metabolites and then spun at 14,000 rcf for 15 min at 4 °C. Four hundred and fifty μL of supernatant was transferred to a new microcentrifuge tube and concentrated overnight using a CentriVap Benchtop Vacuum Concentrator (Labconco, Kanas City, MO, USA). The samples were frozen at −80 °C until metabolomics analysis. he concentrated tissue specimens were thawed, reconstituted in 600 μL of 100 mM sodium phosphate buffer at pH 7.0, and vortexed until the pellets dissolved. The samples were centrifuged at 14,000 rcf for 15 min at 4 °C before transferring 590 μL into 5 mm NMR tubes (Bruker Biospin, USA). Organ: Placenta, Maternal and fetal surfaces |
| Processing Method: | Sonication and solvent removal w/Speed Vac |
| Processing Storage Conditions: | On ice |
| Extraction Method: | 1:1 MeOH/H2O |
| Extract Storage: | -80℃ |
| Sample Resuspension: | 600uL of 100mM sodium phosphate buffer at pH of 7.0 |
Analysis:
| MB Sample ID: | SA069550 |
| Analysis ID: | AN001699 |
| Laboratory Name: | Edison Laboratory |
| Analysis Type: | NMR |
| Software Version: | Topspin 3.6 |
| Operator Name: | Jacquelyn Walejko |
| Num Factors: | 20 |
| Num Metabolites: | 38 |
| Units: | Area under the curve |
NMR:
| NMR ID: | NM000125 |
| Analysis ID: | AN001699 |
| Instrument Name: | Bruker Avance III |
| Instrument Type: | FT-NMR |
| NMR Experiment Type: | 1D 1H |
| Field Frequency Lock: | 2824.58 Hz |
| Spectrometer Frequency: | 600 MHz |
| NMR Solvent: | D2O+H2O |
| NMR Tube Size: | 5mm |
| Shimming Method: | Topshim |
| Pulse Sequence: | cpmgpr1d |
| Water Suppression: | presaturation |
| Power Level: | 5.5584 W |
| Receiver Gain: | 203 |
| Offset Frequency: | 2822.62 Hz |
| Presaturation Power Level: | 2.5963e-05 W |
| Chemical Shift Ref Cpd: | DSS |
| Temperature: | 300 K |
| Number Of Scans: | 128 |
| Dummy Scans: | 16 |
| Acquisition Time: | 1.817 sec |
| Relaxation Delay: | 4 sec |
| Spectral Width: | 15.0207 ppm |
| Num Data Points Acquired: | 32764 |
| Real Data Points: | 65536 |
| Line Broadening: | 0.3 |
| Apodization: | Exponential |
| Baseline Correction Method: | Polynomial order 1 |
| Chemical Shift Ref Std: | 0.00 ppm |
