呼吸道上皮细胞培养模式与纤毛摆动频率的研究进展
|
摘要
呼吸道上皮细胞在呼吸疾病的发生发展中占有重要的地位,哺乳动物呼吸道上皮表面覆盖有纤毛,纤毛的摆动可以保护呼吸系统免于细菌、病毒以及其他致病因素的损害。因此纤毛摆动频率(CBF)的调控成为许多呼吸疾病发生发展的关键因素,可以用来研究呼吸道炎症、吸烟、感染、呼吸系统肿瘤等多种疾病。体外研究CBF的方法多是采用借助呼吸道上皮细胞培养以及活体呼吸道上皮分离等手段,但呼吸道上皮细胞的原代培养具有杂细胞多、不易分化纤毛等诸多问题。呼吸道上皮细胞的培养模式不断发展,本文简述了呼吸道上皮细胞主要的两种原代培养模式及各自的主要培养方法及改进发展情况,同时介绍并总结了呼吸道上皮细胞纤毛分化的鉴定及其摆动频率的检测,明确了各类培养模式对操作流程、培养周期、尤其是纤毛分化及摆动频率的影响,为在现有培养模式下改进技术得到能够诱导纤毛分化的有效培养方法奠定了基础。
Respiratory epithelial cell plays an important role in the development of respiratory diseases. The respiratory epithelial surface of mammalian airway were covered with cilia. The beat of the cilia can protect the respiratory system from bacteria, virus and other harmful factors. Therefore, cilia beat frequency(CBF) became a crucial element of regulating the progress of many kinds of respiratory diseases. It can be used to research inflammation, smoke, infection and respiratory tumor. The technologies of respiratory epithelial cell culture and separation of epithelium in vivo were applied to research CBF. These culture methods, however, had many problems which include low differentiation of cilia and many mixed cells. With the culture mode developed continuously, this paper focused on the major culture mode of respiratory epithelial cells and the culture methods which were improved constantly under relevant culture mode. It also summarized the detection of differentiation of cilia and the CBF, defined the influence on the culture cycle, culture process, especially the differentiation of cilia which were subject to the culture mode, laid the foundation of improving the technology of respiratory epithelial cell culture.
Respiratory epithelial cell plays an important role in the development of respiratory diseases. The respiratory epithelial surface of mammalian airway were covered with cilia. The beat of the cilia can protect the respiratory system from bacteria, virus and other harmful factors. Therefore, cilia beat frequency(CBF) became a crucial element of regulating the progress of many kinds of respiratory diseases. It can be used to research inflammation, smoke, infection and respiratory tumor. The technologies of respiratory epithelial cell culture and separation of epithelium in vivo were applied to research CBF. These culture methods, however, had many problems which include low differentiation of cilia and many mixed cells. With the culture mode developed continuously, this paper focused on the major culture mode of respiratory epithelial cells and the culture methods which were improved constantly under relevant culture mode. It also summarized the detection of differentiation of cilia and the CBF, defined the influence on the culture cycle, culture process, especially the differentiation of cilia which were subject to the culture mode, laid the foundation of improving the technology of respiratory epithelial cell culture.
引文
[1]Wirschell M,Yamamoto R,Alford L,et al.Regulation of ciliary motility:conserved protein kinases and phosphatases are targeted and anchored in the ciliary axoneme[J].Arch Biochem Biophys,2011,510(2):93-100.
[2]Teves ME,Sears PR,Li W,et al.Sperm-associated antigen6(SPAG6)deficiency and defects in ciliogenesis and cilia function:polarity,density,and beat[J].PLo S One,2014,9(10):e107271.
[3]Lee WL,Jayathilake PG,Zhijun T,et al.Muco-ciliary transport:effect of mucus viscosity,cilia beat frequency and cilia density[J].Comp Fluids,2011,49:214-221.
[4]Qin KR,Xiang C.Hysteresis modeling for calcium-mediated ciliary beat frequency in airway epithelial cells[J].Math Biosci,2011,229(1):101-108.
[5]Vetter A,Augustijns P,Bernkop-Schnürch A.Solubilizing agents in nasal formulations and their effect on ciliary beat frequency[J].Toxicol In Vitro,2012,26(1):150-156.
[6]Lee MC,Kim DW,Kim DY,et al.The effect of histamine on ciliary beat frequency in the acute phase of allergic rhinitis[J].Am J Otolaryngol,2011,32(6):517-521.
[7]Bailey KL,Bonasera SJ,Wilderdyke M,et al.Aging causes a slowing in ciliary beat frequency,mediated by PKCε[J].Am J Physiol Lung Cell Mol Physiol,2014,306(6):L854-L859.
[8]Yamaya M,Finkbeiner WE,Chun SY,et al.Differentiated structure and function of cultures from human tracheal epithelium[J].Am J Physiol,1992,262(6 Pt 1):L713-L724.
[9]Davidson DJ,Gray MA,Kilanowski FM,et al.Murine epithelial cells:isolation and culture[J].J Cyst Fibros,2004,Suppl 2:59-62.
[10]Bals R,Beisswenger C,Blouquit S,et al.Isolation and airliquid interface culture of human large airway and bronchiolar epithelial cells[J].J Cyst Fibros,2004,Suppl 2:49-51.
[11]Meng N,Jiao J,Zhang L.Culture of human nasal polyp epithelial cells at an air-liquid surface[J].Chin J Otorhinolaryngol Head Neck Surg,2014,49(1):49-53.
[12]矫健,孟娜,张罗.人鼻腔纤毛上皮细胞的浸没培养[J].临床耳鼻咽喉头颈外科杂志,2014,28(12):898-901.
[13]王达,李煜生.小鼠原代呼吸道上皮细胞分离培养的新方法[J].生理学报,2011,63(6):581-585.
[14]Hirst RA,Jackson CL,Coles JL,et al.Culture of primary ciliary dyskinesia epithelial cells at air-liquid interface can alter ciliary phenotype but remains arobust and informative diagnostic aid[J].PLo S One,2014,9(2):e89675.
[15]Carson JL,Brighton LE,Jaspers I.Phenotypic modification of human airway epithelial cells in air-liquid interface culture induced by exposure to the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK)[J].Ultrastruct Pathol,2015,39(2):104-109.
[16]Dirksen ER,Felix JA,Sanderson MJ.Preparation of explant and organ cultures and single cells from airway epithelium[J].Methods Cell Biol,1995,47:65-74.
[17]宋晓红,张罗,韩德民,等.体外培养的人鼻腔黏膜纤毛的摆动频率[J].中国耳鼻咽喉头颈外科,2010,17(4):175-177.
[18]宋晓红,张罗,韩德民,等.鼠尾胶原为底物的人鼻腔纤毛上皮细胞培养模式的建立[J].中国耳鼻咽喉头颈外科,2007,14(2):107-111.
[19]王奎吉,张罗,韩德民,等.呼吸道纤毛上皮细胞的组织块法培养[J].中国耳鼻咽喉头颈外科,2006,13(12):833-836.
[20]Jiao J,Meng N,Wang H,et al.Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro[J].Front Med,2013,7(4):486-491.
[21]Inoue D,Furubayashi T,Ogawara K,et al.In vitro evaluation of the ciliary beat frequency of the rat nasal epithelium using a high-speed digital imaging system[J].Biol Pharm Bull,2013,36(6):966-973.
[22]Smith CM,Djakow J,Free RC,et al.cilia FA:a research tool for automated,high-throughput measurement of ciliary beat frequency using freely available software[J].Cilia,2012,1:14.
[2]Teves ME,Sears PR,Li W,et al.Sperm-associated antigen6(SPAG6)deficiency and defects in ciliogenesis and cilia function:polarity,density,and beat[J].PLo S One,2014,9(10):e107271.
[3]Lee WL,Jayathilake PG,Zhijun T,et al.Muco-ciliary transport:effect of mucus viscosity,cilia beat frequency and cilia density[J].Comp Fluids,2011,49:214-221.
[4]Qin KR,Xiang C.Hysteresis modeling for calcium-mediated ciliary beat frequency in airway epithelial cells[J].Math Biosci,2011,229(1):101-108.
[5]Vetter A,Augustijns P,Bernkop-Schnürch A.Solubilizing agents in nasal formulations and their effect on ciliary beat frequency[J].Toxicol In Vitro,2012,26(1):150-156.
[6]Lee MC,Kim DW,Kim DY,et al.The effect of histamine on ciliary beat frequency in the acute phase of allergic rhinitis[J].Am J Otolaryngol,2011,32(6):517-521.
[7]Bailey KL,Bonasera SJ,Wilderdyke M,et al.Aging causes a slowing in ciliary beat frequency,mediated by PKCε[J].Am J Physiol Lung Cell Mol Physiol,2014,306(6):L854-L859.
[8]Yamaya M,Finkbeiner WE,Chun SY,et al.Differentiated structure and function of cultures from human tracheal epithelium[J].Am J Physiol,1992,262(6 Pt 1):L713-L724.
[9]Davidson DJ,Gray MA,Kilanowski FM,et al.Murine epithelial cells:isolation and culture[J].J Cyst Fibros,2004,Suppl 2:59-62.
[10]Bals R,Beisswenger C,Blouquit S,et al.Isolation and airliquid interface culture of human large airway and bronchiolar epithelial cells[J].J Cyst Fibros,2004,Suppl 2:49-51.
[11]Meng N,Jiao J,Zhang L.Culture of human nasal polyp epithelial cells at an air-liquid surface[J].Chin J Otorhinolaryngol Head Neck Surg,2014,49(1):49-53.
[12]矫健,孟娜,张罗.人鼻腔纤毛上皮细胞的浸没培养[J].临床耳鼻咽喉头颈外科杂志,2014,28(12):898-901.
[13]王达,李煜生.小鼠原代呼吸道上皮细胞分离培养的新方法[J].生理学报,2011,63(6):581-585.
[14]Hirst RA,Jackson CL,Coles JL,et al.Culture of primary ciliary dyskinesia epithelial cells at air-liquid interface can alter ciliary phenotype but remains arobust and informative diagnostic aid[J].PLo S One,2014,9(2):e89675.
[15]Carson JL,Brighton LE,Jaspers I.Phenotypic modification of human airway epithelial cells in air-liquid interface culture induced by exposure to the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone(NNK)[J].Ultrastruct Pathol,2015,39(2):104-109.
[16]Dirksen ER,Felix JA,Sanderson MJ.Preparation of explant and organ cultures and single cells from airway epithelium[J].Methods Cell Biol,1995,47:65-74.
[17]宋晓红,张罗,韩德民,等.体外培养的人鼻腔黏膜纤毛的摆动频率[J].中国耳鼻咽喉头颈外科,2010,17(4):175-177.
[18]宋晓红,张罗,韩德民,等.鼠尾胶原为底物的人鼻腔纤毛上皮细胞培养模式的建立[J].中国耳鼻咽喉头颈外科,2007,14(2):107-111.
[19]王奎吉,张罗,韩德民,等.呼吸道纤毛上皮细胞的组织块法培养[J].中国耳鼻咽喉头颈外科,2006,13(12):833-836.
[20]Jiao J,Meng N,Wang H,et al.Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro[J].Front Med,2013,7(4):486-491.
[21]Inoue D,Furubayashi T,Ogawara K,et al.In vitro evaluation of the ciliary beat frequency of the rat nasal epithelium using a high-speed digital imaging system[J].Biol Pharm Bull,2013,36(6):966-973.
[22]Smith CM,Djakow J,Free RC,et al.cilia FA:a research tool for automated,high-throughput measurement of ciliary beat frequency using freely available software[J].Cilia,2012,1:14.