Lentivirus was generated by cotransfection of pRRL.SIN-EF1a-PGK-EGFP (encoding green fluorescent protein; #12252) with pRSV-Rev (#12253), pMD2.VSVg (#12259), and pMDL.gag/pol.RRE (#12251) plasmids in 293T cells. After 48 h, viral supernatants were collected, filtered and frozen at −80 °C until use. The LC5 cell line was infected for 24 h with 4 mL of virus-containing medium in presence of 10 µg/mL polybrene. Selection and isolation of fibroblast-GFP+ were performed with Sony Synergy Cell Sorter separation device (Sony Biotechnology Inc., San José, CA, USA) and based on GFP expression and incorporation of Propidium Iodide (PI) for the exclusion of dead cells. All required plasmids were from nonprofit plasmid repository Addgene (http://www.addgene.org/ , accessed on 8 January 2018).
Synergy cell sorter separation device
Manufactured by Sony
Sourced in United States
The Sony Synergy Cell Sorter is a separation device designed for the isolation and purification of specific cell populations from complex biological samples. It utilizes flow cytometry technology to accurately identify and separate target cells based on their unique physical and biochemical properties.
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2 protocols using synergy cell sorter separation device
1
Lentiviral Transduction of LC5 Cells
2
Lentiviral Transduction of LC5 Cells
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Lentivirus was generated by co-transfection of pRRL.SIN-EF1a-PGK-EGFP (encoding green uorescent protein; #12252) with pRSV-Rev (#12253), pMD2.VSVg (#12259), and pMDL.gag/pol.RRE (#12251) plasmids in 293T cells. After 48h, viral supernatants were collected, ltered and frozen at -80 until use.
LC5 cell line was infected for 24h with 4 mL of virus-containing medium in presence of 10 µg/mL polybrene. Selection and isolation of broblasts-GFP + was performed with Sony Synergy Cell Sorter separation device (Sony Biotechnology Inc.), and based of GFP expression, and incorporation of Propidium Iodide (PI) for the exclusion of dead cells. All plasmids required were from non-pro t plasmid repository Addgene (http://www.addgene.org/).
LC5 cell line was infected for 24h with 4 mL of virus-containing medium in presence of 10 µg/mL polybrene. Selection and isolation of broblasts-GFP + was performed with Sony Synergy Cell Sorter separation device (Sony Biotechnology Inc.), and based of GFP expression, and incorporation of Propidium Iodide (PI) for the exclusion of dead cells. All plasmids required were from non-pro t plasmid repository Addgene (http://www.addgene.org/).
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