Culture and differentiation of mouse tracheal epithelial cells at air-liquid interface / 中国药理学通报

Aim Air-liquid interface (ALI) cultures of mouse tracheal epithelial cells (MTEC) are a well-established model to study airway epithelial cells. MTEC provides a powerful ap¬proach for the evaluation of the inhalation toxicological in vitro. Methods C57BL/6 mouse tracheal-bronchial epithelial cells were obtained by digestion with protease in cold temperature o- vemight, and the digestion time was optimized to ensure the quantity and viability of the obtained cells. The cells were cul¬tured into collagen coated Transwell inserts. Proliferating phase and air-liquid interface culture were promoted with different cul¬ture media. The expression of tight junction protein and cell trans-epithelial electrical resistance(TEER) were used to evalu¬ate the formation of tight junction between cells and the analysis of cell polarity. The cilia structure was confirmed by electron mi¬ croscopy and immunofluorescence. Results Highly purified and viable primary airway epithelial cells could be harvested and subcultured by our methods, including morphology and immuno- cytochemistry staining confirmed the expression of MUC5AC, a- tubulin, p-tubulin-IV and ZO-1. The development of tight-junc¬tions and epithelium were similar with pseudostratified ciliated columnar epithelium morphology. Conclusions A comprehen¬sive protocol for ALI culture was established, reproducing the characteristic pseudostratified ciliated columnar epithelium mor¬phology and physiological functions in vitro. The MTEC protocol provides a stable and reliable method for the isolation, mucocili¬ary differentiation and reproducing.

Variable localization of Toll-like receptors in human fallopian tube epithelial cells / 대한생식의학회지

OBJECTIVE: To determine the localization, expression, and function of Toll-like receptors (TLRs) in fallopian tube epithelial cells. METHODS: The localization of TLRs in fallopian tube epithelial cells was investigated by immunostaining. Surprisingly, the intensity of staining was not equal in the secretory and ciliated cells. After primary cell culture of fallopian tube epithelial cells, ring cloning was used to isolate colonies of ciliated epithelial cells, distinct from non-ciliated epithelial cells. The expression of TLRs 1–10 was examined by quantitative real-time polymerase chain reaction, and protein localization was confirmed by immunostaining. The function of the TLRs was determined by interleukin (IL)-6 and IL-8 production in response to TLR2, TLR3, TLR5, TLR7, and TLR9 ligands. RESULTS: Fallopian tube epithelial cells expressed TLRs 1–10 in a cell-type-specific manner. Exposing fallopian tube epithelial cells to TLR2, TLR3, TLR5, TLR7, and TLR9 agonists induced the secretion of proinflammatory cytokines such as IL-6 and IL-8. CONCLUSION: Our findings suggest that TLR expression in the fallopian tubes is cell-type-specific. According to our results, ciliated cells may play more effective role than non-ciliated cells in the innate immune defense of the fallopian tubes, and in interactions with gametes and embryos.

Estudo das emissões otoacústicas durante o ciclo hormonal feminino / Study of otoacoustic emissions during the female hormonal cycle

As alterações hormonais ocorridas em pouco espaço de tempo promovem modificações em todo o organismo da mulher, com manifestações físicas e emocionais muitas vezes evidentes. OBJETIVO: O objetivo deste estudo foi avaliar a atividade das células ciliadas externas em mulheres durante o ciclo menstrual, observando os efeitos das alterações hormonais impostas pelo ciclo em suas três fases. MATERIAL E MÉTODO: Trata-se de um estudo longitudinal prospectivo que avaliou 21 mulheres entre 20 e 35 anos que não faziam uso de medicamento contraceptivo (dados obtidos em anamnese). Foi realizada avaliação das emissões otoacústicas transientes e por produto de distorção durante as três fases do ciclo hormonal (lútea, folicular e ovulatória). Para análise geral dos dados foi utilizado o programa SPSS 13.0, RESULTADOS: As três fases do ciclo hormonal não alteram os valores de amplitude e reprodutibilidade das emissões otoacústicas transientes e por produto de distorção. Foi verificada diferença entre as orelhas na freqüência de 1,5 kHz na amplitude das emissões por produto de distorção, tendo a orelha direita os maiores valores. CONCLUSÃO: Com a realização deste estudo pôde-se concluir que não há diferenças significantes das emissões otoacústicas transientes e por produto de distorção entre as fases do ciclo hormonal.

The hormonal changes that occur in a short time span promote modifications all over the woman’s body, with physical and emotional manifestations which are frequently observed. AIM: to evaluate the activity of the external ciliated cells in women during their menstrual cycle, observing the effect of hormonal changes caused by the cycle in their 3 phases. METODS: this is a longitudinal prospective study where 21 women between 20 and 35 years old who did not take any contraceptive medicine were assessed. Transient otoacoustic emissions were evaluated by distortion product during the 3 phases of the menstrual cycle (luteal, follicular and ovulatory phases). The SPSS 13.0 software was used to analyze the data. RESULTS: the phases of menstrual cycle do not alter the amplitude and reproducibility values of the transient otoacoustic emissions. We noticed a difference between the ears in the frequency of 1.5 KHz in the amplitude of emissions by distortion product, and the right ear showed the highest values. CONCLUSION: There are no significant differences in transient otoacoustic emissions and distortion products in the phases of the menstrual cycle.

La estria supranuclear de las células ciliadas en la rinitis alérgica / Supranuclear stria of ciliated cells in allergic rhinitis

Se estudiaron 80 pacientes adultos alérgicos, que cursaron con los siguientes cuadros clínicos: 16 casos de rinitis intermitente y 64 de rinitis persistente. Se realizó el recuento porcentual de la estría supranuclear de las células ciliadas, respecto de los leucocitos presentes en los extendidos obtenidos por toma endonasal. Con los datos obtenidos se clasificaron los extendidos en 4 grupos; Grupo A (N=23): predominio leucocitario eosinófilo con eosinofilia nasal >10%, Grupo B (N=15): abundantes leucocitos neutrófilos y eosinofilia nasal >10%, Grupo C (N=29): con escasos leucocitos, Grupo D (N=13): con abundantes leucocitos de predominio neutrófilo sin eosinofilia. Se observó que el incremento porcentual de estría supranuclear se correlacionó con eosinofilia nasal >10% y con las muestras que presentaron escasos leucocitos. Sin embargo se evidenció una marcada disminución del porcentaje de estría supranuclear en la leucocitosis neutrófila de etiología bacteriana.

Nasal secretions were studied in 80 allergic adults patients: 16 with intermittent rhinitis and 64 with persistent rhinitis. The percentage of supranuclear stria of ciliated cells with regard to leucocytes was studied by nasal scraping. Four groups of patients were classified according to nasal leucocytic predominance: patients with eosinophilic predominance with eosinophils > 10% in Group A (N=23), patients with abundant neutrophils and eosinophils >10% in Group B (N=15), patients with scant leucocytes in Group C (N=29), patients with neutrophilic predominance without eosinophils in Group D (N=13). An increase of supranuclear stria percentage was correlated to eosinophils > 10% and also correlated to scant leucocytes. Nevertheless, a significant decrease of supranuclear stria percentage was observed in neutrophilic leukocytosis of bacterial etiology.