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比較研究胎牛血清和馬血清對(duì)初級(jí)馬支氣管成纖維細(xì)胞的生長(zhǎng)和分化的影響2016-09-23 13:25
從消化支氣管組織細(xì)胞產(chǎn)量是一致的。在這兩種培養(yǎng)條件,EBF用臺(tái)盼藍(lán)染色和活細(xì)胞的百分率是相似的,通常>95%。下倒光鏡分析,觀察細(xì)胞壞死或凋亡的細(xì)胞沒(méi)有顯著的證據(jù)。對(duì)于細(xì)胞形態(tài),EBF在DMEM中培養(yǎng),用10%FBS的似乎是通常扁平和紡錘形(具有均勻的細(xì)胞質(zhì))在幾個(gè)星期的通道相比EBF在DMEM中培養(yǎng),用10%的HS(圖1的A)。 在FBS的培養(yǎng)基,EBF生長(zhǎng)在生長(zhǎng)表面只有松散的細(xì)胞-細(xì)胞接觸,直到達(dá)到匯合,然后形成了仍然是典型的成纖維細(xì)胞單層,直到通道15道之間EBF緊平行線16-20開(kāi)始改變其形態(tài):細(xì)胞大,平坦和更多邊形的形狀,具有大,異質(zhì)核。同時(shí),細(xì)胞生長(zhǎng)迅速降低和細(xì)胞存活率降低。與此相反,EBF在HS的存在下培養(yǎng)的表現(xiàn)出內(nèi)培養(yǎng)2天改變形態(tài)學(xué)變化;細(xì)胞小,形狀在細(xì)胞質(zhì)顆粒劑般黑暗的結(jié)構(gòu)(圖結(jié)合更緊湊1C)。 此外,細(xì)胞生長(zhǎng)集群和鏈條,未能在一周內(nèi)達(dá)到匯合。EBF的這種形態(tài)行為這種文化條件下看到,直到第7代,但此后,EBF未能定期增殖,并在自己的生存能力(通道9)下降。在這兩種培養(yǎng)條件下,最EBF(>99%)為陽(yáng)性波形蛋白,但與細(xì)胞質(zhì)內(nèi)的培養(yǎng)基含有FBS的(圖更特性絲狀結(jié)構(gòu)1B)的比在HS(圖的存在1D). 總之,血清中除基礎(chǔ)EBF培養(yǎng)基提高EBF分化成肌成纖維細(xì)胞,而這些發(fā)現(xiàn)開(kāi)發(fā)出模擬體內(nèi)的生理過(guò)程,并研究氣道疾病的機(jī)制和重塑體外成纖維細(xì)胞培養(yǎng)模型有用。 英文原文: Comparative study of the effects of fetal bovine serum versus horse serum on growth and differentiation of primary equine bronchial fibroblasts
Primary EBF were cultured on non-collagen coated flasks without serum or in the presence of fetal bovine serum (FBS) or horse serum (HS) or in serum depleted medium. EBF cultured in serum-free basal media and those serum deprived were not able to proliferate and even exhibited considerable cell death. In media containing FBS or HS, proliferation of the cells was reproducible between different primary cultures and cells demonstrated expression of vimentin. Large variations were found in the ability of FBS and HS to support growth and differentiation of EBF in monolayer culture. Indications of growth-promoting actions, increasing passage number as well as maintaining fibroblast morphology were found rather in FBS than in HS. EBF culturing in HS needed longer doubling and confluence time. The protein content of the cell pellets was higher in EBF cultured in medium containing HS than FBS. Alpha-smooth muscle actin seemed to be less expressed in EBF cultured in medium containing FBS than those in HS.
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