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比較研究胎牛血清和馬血清對初級馬支氣管成纖維細胞的生長和分化的影響2016-09-23 13:25
從消化支氣管組織細胞產量是一致的。在這兩種培養條件,EBF用臺盼藍染色和活細胞的百分率是相似的,通常>95%。下倒光鏡分析,觀察細胞壞死或凋亡的細胞沒有顯著的證據。對于細胞形態,EBF在DMEM中培養,用10%FBS的似乎是通常扁平和紡錘形(具有均勻的細胞質)在幾個星期的通道相比EBF在DMEM中培養,用10%的HS(圖1的A)。 在FBS的培養基,EBF生長在生長表面只有松散的細胞-細胞接觸,直到達到匯合,然后形成了仍然是典型的成纖維細胞單層,直到通道15道之間EBF緊平行線16-20開始改變其形態:細胞大,平坦和更多邊形的形狀,具有大,異質核。同時,細胞生長迅速降低和細胞存活率降低。與此相反,EBF在HS的存在下培養的表現出內培養2天改變形態學變化;細胞小,形狀在細胞質顆粒劑般黑暗的結構(圖結合更緊湊1C)。 此外,細胞生長集群和鏈條,未能在一周內達到匯合。EBF的這種形態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。在這兩種培養條件下,最EBF(>99%)為陽性波形蛋白,但與細胞質內的培養基含有FBS的(圖更特性絲狀結構1B)的比在HS(圖的存在1D). 總之,血清中除基礎EBF培養基提高EBF分化成肌成纖維細胞,而這些發現開發出模擬體內的生理過程,并研究氣道疾病的機制和重塑體外成纖維細胞培養模型有用。 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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