Equine yolk sac: a stem cells source / Saco vitelino equino: una fuente de células madre

SUMMARY: Mesenchymal stem cells are characterized by in vitro high proliferation and multilineage potential maintenance. This study aimed to isolate and characterize equine YS mesenchymal stem cells and compare these with amniotic membranes. The yolk sac (YS) and amniotic membranes (AM) were obtained from 20 pregnant mares with gestational age around 30 days. Cells were cultured in α-MEM supplemented with 15 % FBS, 1 % antibiotic solution, 1 % L-glutamine and 1 % nonessential amino acids. To cell characterization we used cytogenetic analysis, fibroblast colony-forming unit assays, cell growth curves, immunophenotyping, flow cytometry, differentiation assays and teratoma formation. Results: Both cell sources presented fibroblastoid and epithelioid-like format. The YS cells have lower colony formation potential then AM ones, 3 versus 8 colonies per 103 plated cells. However, YS cells grew progressively while AM cells showed steady. Both, the YS and amnion cells immunolabeled for Oct-4, Nanog, SSEA-3, cytokeratin 18, PCNA, and vimentin. In addition, presented mesenchymal, hematopoietic, endothelial and pluripotency markers in flow cytometry. Discussion: Both cell sources presented high plasticity and differed into osteogenic, adipogenic, and chondrogenic lineages, and no tumor formation in nude mice was observed. The results suggest that horse YS may be useful for cell therapy such as amnion-derived cells.

RESUMEN: Las células madre mesenquimales se caracterizan por una alta proliferación in vitro y un mantenimiento potencial de múltiples líneas. Este estudio tuvo como objetivo aislar y caracterizar las células madre mesenquimales del saco vitelino equinas y compararlas con las membranas amnióticas. Se obtuvo el saco vitelino (SV) y las membranas amnióticas (MA) de 20 yeguas preñadas con edad gestacional de aproximadamente 30 días. Las células se cultivaron en α -MEM suplementado con 15 % de FBS, 1 % de solución antibiótica, 1 % de L-glutamina y 1 % de aminoácidos no esenciales. Para la caracterización celular utilizamos análisis citogenéticos, ensayos de unidades de colonias de fibroblastos, curvas de crecimiento celular, inmunofenotipaje, citometría de flujo, ensayos de diferenciación y formación de teratomas. Ambas fuentes celulares presentaron formato fibroblastoideo y epitelioide. Las células SV tienen un potencial de formación de colonias más bajo que las de MA, 3 versus 8 colonias por 103 células en placa. Sin embargo, las células SV crecieron progresivamente mientras que las células MA se mostraron estables. Tanto las células YS como las células amnios están inmunomarcadas para Oct-4, Nanog, SSEA-3, citoqueratina 18, PCNA y vimentina. Además, presentó marcadores mesenquimales, hematopoyéticos, endoteliales y pluripotenciales en citometría de flujo. Ambas fuentes celulares presentaron alta plasticidad y diferían en linajes osteogénicos, adipogénicos y condrogénicos, y no se observó formación de tumores en ratones. Los resultados sugieren que el SV de caballo puede ser útil para la terapia celular, como las células derivadas de amnios.

Valoración de enfermería en gestantes con ruptura prematura de membranas / Nursing assessment in pregnant women with premature rupture of membranes

El proyecto se realizó con el objetivo de describir la valoración de Enfermería por dominios según NANDA-Internacional, en gestantes con diagnóstico de RPM, ingresadas en el Servicio de Ginecología y Obstetricia del Hospital Provincial General Docente Riobamba, Ecuador, durante el periodo octubre 2018 - enero 2019. Se realizó un estudio descriptivo, con enfoque cuantitativo, de corte transversal; cuya población de estudio quedó constituida por 61 individuos. Los datos fueron recolectados mediante la aplicación de una entrevista estructurada a cada paciente y de la revisión de documentos (historias clínicas). Se observaron 8 dominios alterados. El 63% de las mujeres participantes tuvo insuficientes controles prenatales. En el 78.7% de los partos se reportó líquido amniótico de aspecto claro. El 73.7% de las gestantes presentó antecedentes de infección. Las mayores afectaciones fueron en los dominios referidos a sexualidad/reproducción y promoción de la salud. La mayoría presentó RPM entre las 39 y 40.6 semanas y la FCF normal. Predominaron las madres adolescentes que declararon no planificar su embarazo.

This research was carried out in order to describe the assessment of Nursing by domains according to NANDA-International in pregnant women with a diagnosis of PROM admitted to the Gynecology and Obstetrics Service of the Provincial General Hospital of Riobamba-Ecuador, during the period October 2018 - January 2019. A cross-sectional and descriptive study was carried out with a quantitative approach; whose study population was made up of 61 individuals. The data were collected by applying a structured interview to each patient and reviewing documents (medical records). 8 altered domains were observed. 63% of the participating women had insufficient prenatal controls. Clear-looking amniotic fluid was reported in 78.7% of deliveries. 73.7% of pregnant women presented a history of infection. The greatest affectations were in the domains referring to sexuality / reproduction and health promotion. The majority presented PROM between 39 and 40.6 weeks and normal FHR. Adolescent mothers who declared not planning their pregnancy predominated.

Fresh Frozen Amniotic Membrane and Buccal Pad of Fat for Reconstruction of Oral Mucosal Defect After Surgical Excision of Leukoplakia

Objective: To compare the use of Fresh Frozen Amniotic Membrane (FFAM) and Buccal Pad of Fat (BPF) for reconstruction of oral mucosal defect after surgical excision of leukoplakia. Material and Methods: Twenty patients were randomly selected and divided into two groups. Group 1 use amniotic membrane graft and Group 2 BPF. Both groups were evaluated preoperatively and postoperatively. Incisal opening, epithelialization and fibrosis were evaluated after one month of surgery. Chi square and Student t tests were used. Results: According to the presence of smoking habits, the highest frequencies were for smoking (30%) and betel leaf areca nut with tobacco (30%). Regarding the diameter of oral leukoplakia, in 40% of the participants it was 2x3 cm2. In Group1, after one month of surgery preoperative and postoperative inter-incisal opening values were 44.20 ± 3.37 and 42.05 ± 3.47 (p<0.001). In Group 2, preoperative and postoperative inter-incisal opening values were 44.09 ± 3.32 and 43.01±3.38 (p>0.05). When FFAM was used complete epithelialization in 70% and incomplete epithelialization in 30% patients. When BPF was used the results were almost similar. Fibrosis occurred in 30% in Group 1. There were no complications like flap necrosis, infectiona and hematoma formation. Conclusion: Incisal opening was significantly better in Fresh Frozen Amniotic Membrane Group, epithelialization and fibrosis were almost same in both groups after surgical excision of oral leukoplakia.

Transcription factor Ets-1 expression in human fetal membranes and its clinical significance / 中华妇产科杂志

Objective To investigate the role of transcription factor Ets-1 involved in premature rupture of membranes(PROM) by detecting its expression and location in human fetal membranes. Methods Between Feb. and Nov. 2007, 100 pregnant women who delivered in the First Affiliated Hospital, Chongqing Medical University were enrolled in this study. According to gestational weeks, rupture of amuiochorionic membranes and delivery mode, those women were classified into preterm labor group, preterm premature rupture of membranes (PPROM) group, term labor group and term premature rupture of membranes (TPROM) group, matched with elective term cesarean sections women as control group. There were 20 pregnant women in every group. The expression and distribution of Ets-1 protein in fetal membranes were detected by immunohistochemical streptravidin-biotin peroxidase(SP) method and histoscore. In the mean time, 6 cases were chosen from each group randomly and reverse transcription-PCR was used to measure the Ets-1 mRNA expression. Results (1) The expression of Ets-1 mRNA in fetal membranes were 0.342±0.016 in preterm labor group,0.603±0.027 in PPROM group,0.325±0.013 in term labor group, 0.582±0.075 in TPROM group,0.139±0.012 in control group, respectively. When compared with that in control group, Ets-1 mRNA expression were significantly increased in both PPROM and TPROM group(P< 0.05). However, it did not show remarkably difference between preterm group and term labor group, as well as between PPROM and TPROM group (P>0.05). (2) Ets-1 was in stromal layers of amniochorionic membranes and in both cytoplasm and nuclei of trophoblast, which were shown with diffuse intracellular positive granularities (brown-yellow) clearly. No expression of Ets-1 was observed in amniotic epithelial cells. (3) The expression of Ets-1 protein was 0.552±0.018 in preterm labor group, 2.853±0.174 in PPROM group, 0.538±0.042 in term labor group, 2.731±0.090 in TPROM group and 0.214±0.013 in control group, respectively. Ets-1 protein was significantly increased in the stroma of both PPROM and TPROM group than that in control group(P<0.05). However, no remarkable different expression of Ets-1 was observed between preterm and term labor group,so was that between PPROM and TPROM group(P> 0.05). Conclusion Transcription factor Ets-1 is expressed in human amniochorionic membranes and it could be up-regulated in PROM.

Expression of aquaporin 8 in human fetal membrane and placenta of idiopathic polyhydramnios / 中华妇产科杂志

Objective To determine the expression of Aquaporin 8 (AQP8) in the fetal membrane and placenta of idiopathic polyhydramnios. Methods The amnion, chorion and placenta were collected from 12 term pregnancies with idiopathic polyhydramnios(polyhydramnios group) and 12 term pregnancies who were normal (control group). The expression of AQP8 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). The expression of AQP8 protein was detected by immunohistochemistry. Results The expression of AQP8 mRNA in amnion, chorion and placenta of polyhydramnios group was (0.78±0.13), (0.58±0.10), and (0.86±0.15) respectively, and that of control group was (0.39±0.07 ), (0.45±0.09),and (0.34±0.09) respectively. The expression of AQP8 protein in amnion, chorion and placenta of polyhydramnios group was (0.195±0.024), (0. 170±0.028), and (0. 193±0.024) respectively, and that of control group was (0. 151±0.018), (0.156±0.024), and (0. 152±0.023) respectively. In all 3 types of tissues the expression of AQP8 mRNA of polyhydramnios group was higher than that of control group (P<0.05). In anmion and placenta the expression of AQP8 protein of polyhydramnios group was also increased compared to that of control group (P<0.05), but in chorion the difference in AQP8 protein expression between the two groups was not significant(P>0.05). Conclusion The expression of AQP8 mRNA and protein is significantly increased in the anmion and placenta of polyhydramnios, suggesting that AQP8 may play an important role in the regulation of amniotic fluid volume.

Expression of aquaporin-1,3,8,9 mRNA in human amniotic membranes in polyhydranmios / 中华围产医学杂志

Objective To determine the expression of aquaporin-1,3,8,9 mRNA (AQP-1,3,8, 9) in amniotic membranes in pregnant women with polyhydramnios. Methods Amniotic membranes were collected from women who presented with either polyhydramnios (n= 5)or normal amniotic fluid volume (control, n= 5) underwent elective cesarean sections at term. The AQP-1,3,8,9 mRNA expression were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Results The expression of AQP-9 mRNA on fetal membranes were significantly higher in polyhydramnios groups (1. 1403±0. 0831) than that of control (0. 5903±0. 1909) (P = 0. 002), although the expression of AQP-1, 3 and 8 showed no significant difference between the two groups (P= 0. 972, 0. 242,0. 608, respectively). Conclusions AQP-9 may play an important role in maintaining the volume and the balance of different components of amniotic fluid in polyhydramnios cases.

Expression of aquaporin 3 and aquaporin 9 in placenta and fetal membrane with idiopathic polyhydramnios / 中华妇产科杂志

Objective To investigate the pathogenesis role of aquaporin 3 and aquaporin 9 in idiopathic polyhydramnios by detecting their expression and distribution in fetal membranes and placenta.Methods Twenty-one of term pregnancy women with idiopathic polyhydramnios were enrolled as patient group matched with 30 women with normal term pregnancy as control group.The expression and localization of aquaporin 3 and aquaporin 9 in fetal membranes and placenta were detected by real-time polymerase chain reaction and streptavidin peroxidase immunohistochemiscal staining.Results (1)The mRNA expressions of aquaporin 3 and aquaporin 9 were detected in amnion,chorion and placental tissue in both patient group and control group.Both aquaporin 3 and aquaporin 9 were demonstrated positive staining in the amnion epithelia,chorion cytotrophoblasts and placental trophoblast.(2)The ratio of aquaporin 3 and aquaporin 9 mRNA expressions in amnion in patient group comparing to those in control group were 5.00 and 3.25,while in chorion they were 2.03 and 2.08.When compared with those in amnion and chorion of control group,there was a significant difference(P<0.01).However,the relative change fold of aquaporin 3 and aquaporin 9 in placental trophoblast in patient group were decreased in comparison of those in control group,which also showed statistical difference(P<0.01).(3)The expression of aquaporin 3 and aquaporin 9 protein in anmion were 7.5 ±2. 0 and 11.1 ± 1.8 in patient group, while they were 5.3 ± 1. 6 and 5.6 ± 2. 3 in control group. In chorion, the expression of aquaporin 3 and aquaporin 9 protein was 7.5±2. 0 and 10. 0 ±1.6 in patient group, respectively, while in control group, they were 5.4 ±2.2 and 5.6±2. 1. When compared with those proteins in control group, it exhibited statistical difference (P<0.05). However, in placental trophoblast of patient greup,the expression of aquaporin 3 and aquaporin 9 protein were 3.5±1.4and 4. 0±2. 5, respectively, which were significantly decreased than 5.6±1. 3 and 7. 1±2. 9 in control group(P< 0. 05). Conclusions The alterations of aquaporin 3 and aquaporin 9 expressions in fetal membrane and placenta might be an adaptive response to idiopathic polyhydramnios. Further investigation should be needed to clarify the regulatory mechanism of aquaporin 3 and aquaporin 9 expressions.