![]() The RNA samples were treated with DNase I (Sigma-Aldrich, St Louis, USA), quantified, and reverse-transcribed into cDNA with the ReverTra Ace- First Strand cDNA Synthesis Kit (TOYOBO, TOYOBO (SHANGHAI) BIOTECH CO., LTD., Shanghai, China). Total RNA from each cell was isolated with Trizol Reagent (Invitrogen, Life Technologies Corporation, Grand Island, NY, USA), according to the manufacturer's protocol. In this experiment, all cells had been cultured on the same conditions until passage 4th before making ulterior experiments. All CD44+CD105+ cells were cultured in a humidified incubator, at 37℃ with 5%CO2, until 80% confluent. The subpopulation cells were plated at 1×106 cells/ml in DMEM:F12 (1:1) (Gibco, Gaithersburg, MD, USA), supplemented with 10ng/ml basic fibroblast growth factor (bFGF), 10ng/ml epidermal growth factor (EGF) (all from Sigma-Aldrich, St Louis, USA), 10% fetal bovine serum and 2mM L-glutamine (all from Gibco, Gaithersburg, MD, USA). After reaction, the cells were washed twice in PBS, and were put the secondary monoclonal antibodies (Goat anti-mouse or Goat anti-rabbit coupled to magnetic microbeads, Miltenyi Biotec, Auburn, CA), incubated at 10℃ in PBS for 15 min and then washed twice in PBS. CD44+/CD105+ subpopulation cells were isolated from those human amniotic fluid using 4μl of the primary monoclonal antibodies (mouse anti-human CD44-FITC, rabbit anti-human CD105-PE, eBioscience Inc, San Diego, CA, USA)) stored at 4℃ in PBS for 30 min in a volume of 1ml as previously described. All of the 10 human samples were obtained after approval from the Ethical Review Board of the Shanghai First Maternity and Infant Hospital (Shanghai, China) and after obtaining written informed consent from subjects. doi:10.3109/ amniotic fluids were obtained by ultrasound-guided amniocentesis performed on pregnant women for routine prenatal diagnosis purposes at gestational ages ranging from 18 th -22th weeks. Amniotic Fluid Sludge as a Marker of Intra-Amniotic Infection and Histological Chorioamnionitis in Cervical Insufficiency: A Report of Four Cases and Literature Review. Paules C, Moreno E, Gonzales A, Fabre E, González de Agüero R, Oros D. Effect of Antibiotic Treatment of Amniotic Fluid Sludge. Presence of Amniotic Fluid Sludge and Pregnancy Outcomes: A Systematic Review. Pergialiotis V, Bellos I, Antsaklis A, Loutradis D, Daskalakis G. Antibiotic Treatment of Amniotic Fluid "Sludge" in Patients During the Second or Third Trimester with Uterine Contraction. Jin W, Ha Kim Y, Kim J, Kim T, Kim A, Yang Y. ![]() ![]() Ultrasound Differential Diagnosis Between Amniotic Fluid Sludge and Blood Clot from Placental Edge Separation. Kantorowska A, Kunzier N, Kidd J, Vintzileos A. What is Amniotic Fluid 'Sludge'? Ultrasound Obstet Gynecol. It is proposed that amniotic fluid sludge is an indicator of intra-amniotic microbial invasion correlating with histological chorioamnionitis and funisitis 6. It is also usually associated with a shortened cervical length when the patient presents with both aspects, the pre-term labor risk increases 5. Amniotic fluid sludge is associated with spontaneous pre-term labor and, therefore, lower gestational age at birth, lower neonatal birthweight, and increased neonatal intensive care unit admission rate and neonatal death 4. Clinical significanceĪmniotic fluid sludge has been identified as a potential marker of adverse pregnancy and perinatal outcomes. ![]() On aspiration, amniotic fluid sludge is usually cloudy and thick with an elevated white cell count and may be due to a microbial invasion of the amniotic cavity 1. Amniotic fluid sludge represents a collection of bacterial and inflammatory cells embedded in an amorphous material consistent with microbial biofilm 3. ![]()
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