期刊论文详细信息
BMC Biotechnology
Use of polyethyleneimine polymer in cell culture as attachment factor and lipofection enhancer
Rafael P Ballestero1  Maribel González-García1  Madhuri Jasti1  Kishore VL Parsa1  Suman Govindaraju1  Ajith R Vancha1 
[1]Departments of Chemistry and Biology, Texas A&M University-Kingsville, Kingsville, TX 78363, USA
Others  :  1154516
DOI  :  10.1186/1472-6750-4-23
 received in 2004-05-15, accepted in 2004-10-15,  发布年份 2004
PDF
【 摘 要 】

Background

Several cell lines and primary cultures benefit from the use of positively charged extracellular matrix proteins or polymers that enhance their ability to attach to culture plates. Polyethyleneimine is a positively charged polymer that has gained recent attention as a transfection reagent. A less known use of this cationic polymer as an attachment factor was explored with several cell lines.

Results

Polyethyleneimine compared favorably to traditional attachment factors such as collagen and polylysine. PC-12 and HEK-293 cells plated on dishes coated with polyethyleneimine showed a homogeneous distribution of cells in the plate, demonstrating strong cell adhesion that survived washing procedures. The polymer could also be used to enhance the adherence and allow axonal outgrowth from zebrafish retinal explants. The effects of this coating agent on the transfection of loosely attaching cell lines were studied. Pre-coating with polyethyleneimine had the effect of enhancing the transfection yield in procedures using lipofection reagents.

Conclusion

Polyethyleneimine is an effective attachment factor for weakly anchoring cell lines and primary cells. Its use in lipofection protocols makes the procedures more reliable and increases the yield of expressed products with commonly used cell lines such as PC-12 and HEK-293 cells.

【 授权许可】

   
2004 Vancha et al; licensee BioMed Central Ltd.

【 预 览 】
附件列表
Files Size Format View
20150407104845310.pdf 2504KB PDF download
Figure 5. 39KB Image download
Figure 4. 31KB Image download
Figure 3. 35KB Image download
Figure 2. 53KB Image download
Figure 1. 73KB Image download
【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】
  • [1]Kleinman HK, Klebe RJ, Martin GR: Role of collagenous matrices in the adhesion and growth of cells. J Cell Biol 1981, 88:473-485.
  • [2]Kleinman HK, Luckenbill-Edds L, Cannon FW, Sephel GC: Use of extracellular matrix components for cell culture. Anal Biochem 1987, 166:1-13.
  • [3]Yavin E, Yavin Z: Attachment and culture of dissociated cells from rat embryo cerebral hemispheres on polylysine-coated surface. J Cell Biol 1974, 62:540-546.
  • [4]Letourneau PC: Possible roles for cell-to-substratum adhesion in neuronal morphogenesis. Dev Biol 1975, 44:77-91.
  • [5]Boussif O, Lezoualc'h F, Zanta MA, Mergny MD, Scherman D, Demeneix B, Behr JP: A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci U S A 1995, 92:7297-7301.
  • [6]Boussif O, Zanta MA, Behr JP: Optimized galenics improve in vitro gene transfer with cationic molecules up to 1000-fold. Gene Ther 1996, 3:1074-1080.
  • [7]Kircheis R, Kichler A, Wallner G, Kursa M, Ogris M, Felzmann T, Buchberger M, Wagner E: Coupling of cell-binding ligands to polyethylenimine for targeted gene delivery. Gene Ther 1997, 4:409-418.
  • [8]Horbinski C, Stachowiak MK, Higgins D, Finnegan SG: Polyethyleneimine-mediated transfection of cultured postmitotic neurons from rat sympathetic ganglia and adult human retina. BMC Neurosci 2001, 2:2. BioMed Central Full Text
  • [9]Ogris M, Steinlein P, Carotta S, Brunner S, Wagner E: DNA/polyethylenimine transfection particles: influence of ligands, polymer size, and PEGylation on internalization and gene expression. AAPS PharmSci 2001, 3:E21.
  • [10]Guerra-Crespo M, Charli JL, Rosales-Garcia VH, Pedraza-Alva G, Perez-Martinez L: Polyethylenimine improves the transfection efficiency of primary cultures of post-mitotic rat fetal hypothalamic neurons. J Neurosci Methods 2003, 127:179-192.
  • [11]Simon KO, Nutt EM, Abraham DG, Rodan GA, Duong LT: The alphavbeta3 integrin regulates alpha5beta1-mediated cell migration toward fibronectin. J Biol Chem 1997, 272:29380-29389.
  • [12]Robbins AK, Horlick RA: Macrophage scavenger receptor confers an adherent phenotype to cells in culture. Biotechniques 1998, 25:240-244.
  • [13]Turner DC, Flier LA, Carbonetto S: Identification of a cell-surface protein involved in PC12 cell-substratum adhesion and neurite outgrowth on laminin and collagen. J Neurosci 1989, 9:3287-3296.
  • [14]Dwyer DS, Liu Y, Bradley RJ: An ethanol-sensitive variant of the PC12 neuronal cell line: sensitivity to alcohol is associated with increased cell adhesion and decreased glucose accumulation. J Cell Physiol 1999, 178:93-101.
  • [15]Ruegg UT, Hefti F: Growth of dissociated neurons in culture dishes coated with synthetic polymeric amines. Neurosci Lett 1984, 49:319-324.
  • [16]Lelong IH, Petegnief V, Rebel G: Neuronal cells mature faster on polyethyleneimine coated plates than on polylysine coated plates. J Neurosci Res 1992, 32:562-568.
  • [17]Bledi Y, Domb AJ, Linial M: Culturing neuronal cells on surfaces coated by a novel polyethyleneimine-based polymer. Brain Res Brain Res Protoc 2000, 5:282-289.
  • [18]Greene LA, Tischler AS: Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc Natl Acad Sci U S A 1976, 73:2424-2428.
  • [19]Zhuo M, Yu FR, Xu DH, Sun LY, Liu XY: Baculovirus p35 gene greatly enhances PC12 cell's resistance against oxidative stress. J Neurol Sci 2003, 216:135-141.
  • [20]Marszalek JR, Kitidis C, Dararutana A, Lodish HF: Acyl CoA synthetase 2 (ACS2) over-expression enhances fatty acid internalization and neurite outgrowth. J Biol Chem 2004.
  • [21]Landreth GE, Agranoff BW: Explant culture of adult goldfish retina: effect of prior nerve crush. Brain Res 1976, 118:299-303.
  • [22]Agranoff BW, Ford-Holevinski TS: Biochemical aspects of the regenerating goldfish visual system. In Axonal transport in neuronal growth and regeneration. Edited by Elam JS and Cancalon P. New York, Plenum Press; 1984:69-86.
  • [23]Zhang Z, Baron R, Horne WC: Integrin engagement, the actin cytoskeleton, and c-Src are required for the calcitonin-induced tyrosine phosphorylation of paxillin and HEF1, but not for calcitonin-induced Erk1/2 phosphorylation. J Biol Chem 2000, 275:37219-37223.
  • [24]Kosswig N, Rice S, Daugherty A, Post SR: Class A scavenger receptor-mediated adhesion and internalization require distinct cytoplasmic domains. J Biol Chem 2003, 278:34219-34225.
  • [25]Ballestero RP, Dybowski JA, Levy G, Agranoff BW, Uhler MD: Cloning and characterization of zRICH, a 2',3'-cyclic-nucleotide 3'- phosphodiesterase induced during zebrafish optic nerve regeneration. J Neurochem 1999, 72:1362-1371.
  • [26]Fiennes AG, Walton J, Winterbourne D, McGlashan D, Hermon-Taylor J: Quantitative correlation of neutral red dye uptake with cell number in human cancer cell cultures. Cell Biol Int Rep 1987, 11:373-378.
  • [27]Inohara N, Gourley TS, Carrio R, Muniz M, Merino J, Garcia I, Koseki T, Hu Y, Chen S, Nunez G: Diva, a Bcl-2 homologue that binds directly to Apaf-1 and induces BH3-independent cell death. J Biol Chem 1998, 273:32479-32486.
  • [28]Sambrook J, Fritsch EF, Maniatis T: Molecular cloning. A laboratory manual. Second edition. New York, Cold Spring Harbor Laboratory Press; 1989.
  文献评价指标  
  下载次数:40次 浏览次数:4次