Pluronic PF68 increases transfection efficiency in electroporationof mesenchymal stem cells

Pluronic PF68 increases transfection efficiency in electroporationof mesenchymal stem cells

Electroporation is a safe and efficient method to transfect human tooth germ stem cells (hTGSCs), which are mainly mesenchymal stem cells (MSCs). In this study, pluronic F68 (PF68), a nonionic and low-foaming surfactant, was used for the first time to increase the transfection efficiency in electroporation of hTGSCs. Our results revealed that when PF68 was used in the preincubation, electroporation, and recovery medium, it dramatically increased transfection efficiency. Moreover, use of PF68 did not alter the differentiation capacity of hTGSCs, whereas cells electroporated without PF68 had significantly lower expression of Oct-4, pluripotency marker gene, and alkaline phosphatase (ALP) activity upon osteogenic differentiation. We found that PF68 increases transfection efficiency in both electroporation and chemical transfection of cancer cells lines MCF-7 and HeLa. The study suggested that PF68 might be a safe and reliable chemical to use for increasing transfection efficiency during electroporation and chemical transfections.

___

  • ReferencesAyuni EL, Gazdhar A, Giraud MN, Kadner A, Gugger M, Cecchini M, Caus T, Carrel TP, Schmid RA, Tevaearai HT (2010). In vivo electroporation mediated gene delivery to the beating heart. PLoS One 5: e14467.
  • Campagnoli C, Roberts IA, Kumar S, Bennett PR, Bellantuono I, Fisk NM (2001). Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone mar-row. Blood 98: 2396-2402.
  • Collins JM, Despa F, Lee RC (2007). Structural and functional recov-ery of electropermeabilized skeletal muscle in-vivo after treat-ment with surfactant poloxamer 188. Biochim Biophys Acta 1768: 1238-1246.
  • Dogan A, Yalvac ME, Sahin F, Kabanov AV, Palotas A, Rizvanov AA (2012). Differentiation of human stem cells is promoted by amphiphilic pluronic block copolymers. Int J Nanomedicine 7: 4849-4860.
  • Dogan A, Yalvac ME, Yilmaz A, Rizvanov A, Sahin F (2013). Effect of F68 on cryopreservation of mesenchymal stem cells derived from human tooth germ. Appl Biochem Biotechnol 171: 1819-1831.
  • Ferreira E, Potier E, Logeart-Avramoglou D, Salomskaite-Davalgiene S, Mir LM, Petite H (2008). Optimization of a gene electro-transfer method for mesenchymal stem cell transfection. Gene Ther 15: 537-544.
  • Gehl J (2003). Electroporation: theory and methods, perspectives for drug delivery, gene therapy and research. Acta Physiol Scand 177: 437-447.
  • Graziano A, d’Aquino R, Laino G, Papaccio G (2008). Dental pulp stem cells: a promising tool for bone regeneration. Stem Cell Rev 4: 21-26.
  • Hartikka J, Sukhu L, Buchner C, Hazard D, Bozoukova V, Margalith M, Nishioka WK, Wheeler CJ, Manthorp M, Sawdey M (2001). Electroporation-facilitated delivery of plasmid DNA in skeletal muscle: plasmid dependence of muscle damage and effect of poloxamer 188. Mol Ther 4: 407-415.
  • Ikeda E, Yagi K, Kojima M, Yagyuu T, Ohshima A, Sobajima S, Tado-koro M, Katsube Y, Isoda K, Kondoh M et al. (2008). Multipo-tent cells from the human third molar: feasibility of cell-based therapy for liver disease. Differentiation 76: 495-505.
  • Peister A, Mellad JA, Wang M, Tucker HA, Prockop DJ (2004). Stable transfection of MSCs by electroporation. Gene Ther 11: 224-228.
  • Sharma V, Stebe K, Murphy JC, Tung L (1996). Poloxamer 188 de-creases susceptibility of artificial lipid membranes to electro-poration. Biophys J 71: 3229-3241.
  • Tasli PN, Yalvac ME, Sofiev N, Sahin F (2013). Effect of F68, F127, and P85 pluronic block copolymers on odontogenic differen-tiation of human tooth germ stem cells. J Endod 39: 1265-1271.
  • Tung L, Troiano GC, Sharma V, Raphael RM, Stebe KJ (1999). Changes in electroporation thresholds of lipid membranes by surfactants and peptides. Ann NY Acad Sci 888: 249-265.
  • Yalvac ME, Ramazanoglu M, Gumru OZ, Sahin F, Palotas A, Rizva-nov AA (2009). Comparison and optimisation of transfection of human dental follicle cells, a novel source of stem cells, with different chemical methods and electro-poration. Neurochem Res 34: 1272-1277.
  • Yalvac ME, Ramazanoglu M, Rizvanov AA, Sahin F, Bayrak OF, Salli U, Palotas A, Kose GT (2010). Isolation and characterization of stem cells derived from human third molar tooth germs of young adults: implications in neo-vascularization, osteo-, adi-po- and neurogenesis. Pharmacogenomics J 10: 105-113.
  • Yalvac ME, Rizvanov AA, Kilic E, Sahin F, Mukhamedyarov MA, Is-lamov RR, Palotas A (2009). Potential role of dental stem cells in the cellular therapy of cerebral ischemia. Curr Pharm Des 15: 3908-3916.Yalvac ME, Yarat A, Mercan D, Rizvanov AA, Palotas A, Sahin F (2013). Characterization of the secretome of human tooth germ stem cells (hTGSCs) reveals neuro-protection by fine-tuning micro-environment. Brain Behav Immun 32: 122-130.
  • Yalvac ME, Yilmaz A, Mercan D, Aydin S, Dogan A, Arslan A, Demir Z, Salafutdinov II, Shafigullina AK, Sahin F et al. (2011). Dif-ferentiation and neuro-protective properties of immortalized human tooth germ stem cells. Neurochem Res 36: 2227-2235.
  • Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, Al-fonso ZC, Fraser JK, Benhaim P, Hedrick MH (2002). Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13: 4279-4295.
Turkish Journal of Biology-Cover
  • ISSN: 1300-0152
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK
Sayıdaki Diğer Makaleler

Jayabalan SHILPHA, Manoharan JAYASHRE, Muthiah Joe Virgin LARGIA, Manikandan RAMESH

Purification and properties of lipoxygenase from opium poppy seedlings(Papaver somniferum L.)

Ivana HOLKOVA, Frantısek BILKA, Drohomira RAUOVA, Lydia BEZAKOVA

Rejuvenation of mature lentisk by micrografting and evaluation of genetic stability

Ahu UNCUOĞLU ALTINKUT, Muhammet ŞAKİROĞLU, Engin TİLKAT, Hakan YILDIRIM, Yelda ÇİFTÇİ ÖZDEN, İbrahim KOÇ, Fatih Mehmet KILINÇ, Özge METİN KAAKAŞ, Nazan ÇALAR, Ömer Faruk AKDEMİR, Ahmet ONAY

Advancement in protocol for in vitro seed germination, regeneration, bulblet maturation, and acclimatization of Fritillaria persica

Cengiz SANCAK, Derya ÇAKMAK, Cuma KARAOĞLU, Sebahatin ÖZCAN, Muhammed A. ASIM

Genome-wide analysis of response to low sulfur (LSU) genes in grass species and expression profiling of model grass species Brachypodium distachyon under S deficiency

Ertuğrul FİLİZ, Hüseyin TOMBULOĞLU, Abdugaffar ABLAZOV

Prokaryotic expression and purification of porcine Sox6

Wanxe WEN, Jungio LUO, Ping ZHENG, Jie YU, Meng XU, Xingbing MAO, Jun HE, Xiaoling CHEN, Zhiqing HUANG, Daiwen CHEN, Bing YU

İvana HOLKOVÁ, Frantisek BILKA, Drahomíra RAUOVÁ, Lydia BEZÁKOVÁ

Prajitha VAZHANGAT, John Ernest THOPPIL

Hanife GENÇ, Marc F. SCHETELIG, Xavier NIRMALA, Alfred M. HANDLER

Apoptotic induction via membrane/DNA damage and metabolic inactivation by synthetic food colorants in Allium cepa root meristem

Prajitha VAZHANGAT, John Ernest THOPPIL