Cloning, Expression, and Functional Characterization of In-House Prepared Human Leukemia Inhibitory Factor
Objective: Leukemia inhibitory factor (LIF) plays important roles in cellular proliferation, growth promotion and differentiation of various types of target cells. In addition, LIF influences bone metabolism, cachexia, neural development, embryogenesis and inflammation. Human LIF (hLIF) is an essent...
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| Format: | Article |
| Language: | English |
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Royan Institute (ACECR), Tehran
2013-01-01
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| Series: | Cell Journal |
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| Online Access: | http://celljournal.org/library/upload/article/af_628374225443472322342324262733245325552213-Rassouli.pdf |
| _version_ | 1818925544305065984 |
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| author | Hassan Rassouli Shiva Nemati Siamak Rezaeiani Ali Sayadmanesh Mohammad Reza Gharaati Ghasem Hosseini Salekdeh Hossein Baharvand Hamid Gourabi |
| author_facet | Hassan Rassouli Shiva Nemati Siamak Rezaeiani Ali Sayadmanesh Mohammad Reza Gharaati Ghasem Hosseini Salekdeh Hossein Baharvand Hamid Gourabi |
| author_sort | Hassan Rassouli |
| collection | DOAJ |
| description | Objective: Leukemia inhibitory factor (LIF) plays important roles in cellular proliferation, growth promotion and differentiation of various types of target cells. In addition, LIF influences bone metabolism, cachexia, neural development, embryogenesis and inflammation. Human LIF (hLIF) is an essential growth factor for the maintenance of mouse embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in a pluripotent, undifferentiated state.Materials and Methods: In this experimental study, we cloned hLIF into the pENTR-D/TOPO entry vector by a TOPO reaction. Next, hLIF was subcloned into the pDEST17 destination vector by the LR reaction, which resulted in the production of a construct that was transferred into E. coli strain Rosetta-gami™ 2(DE3) pLacI competent cells to produce the His6-hLIF fusion protein.Results: This straightforward method produced a biologically active recombinant hLIF protein in E. coli that has long-term storage ability. This procedure has provided rapid, cost effective purification of a soluble hLIF protein that is biologically active and functional as measured in mouse ESCs and iPSCs in vitro.Conclusion: Our results showed no significant differences in function between laboratory produced and commercialized hLIF. |
| first_indexed | 2024-12-20T02:42:55Z |
| format | Article |
| id | doaj.art-a816235b382f4a909f4520d4901a1a11 |
| institution | Directory Open Access Journal |
| issn | 2228-5806 2228-5814 |
| language | English |
| last_indexed | 2024-12-20T02:42:55Z |
| publishDate | 2013-01-01 |
| publisher | Royan Institute (ACECR), Tehran |
| record_format | Article |
| series | Cell Journal |
| spelling | doaj.art-a816235b382f4a909f4520d4901a1a112022-12-21T19:56:16ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142013-01-01152190197Cloning, Expression, and Functional Characterization of In-House Prepared Human Leukemia Inhibitory FactorHassan RassouliShiva NematiSiamak RezaeianiAli SayadmaneshMohammad Reza GharaatiGhasem Hosseini SalekdehHossein BaharvandHamid GourabiObjective: Leukemia inhibitory factor (LIF) plays important roles in cellular proliferation, growth promotion and differentiation of various types of target cells. In addition, LIF influences bone metabolism, cachexia, neural development, embryogenesis and inflammation. Human LIF (hLIF) is an essential growth factor for the maintenance of mouse embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in a pluripotent, undifferentiated state.Materials and Methods: In this experimental study, we cloned hLIF into the pENTR-D/TOPO entry vector by a TOPO reaction. Next, hLIF was subcloned into the pDEST17 destination vector by the LR reaction, which resulted in the production of a construct that was transferred into E. coli strain Rosetta-gami™ 2(DE3) pLacI competent cells to produce the His6-hLIF fusion protein.Results: This straightforward method produced a biologically active recombinant hLIF protein in E. coli that has long-term storage ability. This procedure has provided rapid, cost effective purification of a soluble hLIF protein that is biologically active and functional as measured in mouse ESCs and iPSCs in vitro.Conclusion: Our results showed no significant differences in function between laboratory produced and commercialized hLIF.http://celljournal.org/library/upload/article/af_628374225443472322342324262733245325552213-Rassouli.pdfLeukemia Inhibitory FactorRecombinant ProteinEmbryonic Stem CellsInduced Pluripotent Stem CellsCell Proliferation |
| spellingShingle | Hassan Rassouli Shiva Nemati Siamak Rezaeiani Ali Sayadmanesh Mohammad Reza Gharaati Ghasem Hosseini Salekdeh Hossein Baharvand Hamid Gourabi Cloning, Expression, and Functional Characterization of In-House Prepared Human Leukemia Inhibitory Factor Cell Journal Leukemia Inhibitory Factor Recombinant Protein Embryonic Stem Cells Induced Pluripotent Stem Cells Cell Proliferation |
| title | Cloning, Expression, and Functional Characterization of In-House Prepared Human Leukemia Inhibitory Factor |
| title_full | Cloning, Expression, and Functional Characterization of In-House Prepared Human Leukemia Inhibitory Factor |
| title_fullStr | Cloning, Expression, and Functional Characterization of In-House Prepared Human Leukemia Inhibitory Factor |
| title_full_unstemmed | Cloning, Expression, and Functional Characterization of In-House Prepared Human Leukemia Inhibitory Factor |
| title_short | Cloning, Expression, and Functional Characterization of In-House Prepared Human Leukemia Inhibitory Factor |
| title_sort | cloning expression and functional characterization of in house prepared human leukemia inhibitory factor |
| topic | Leukemia Inhibitory Factor Recombinant Protein Embryonic Stem Cells Induced Pluripotent Stem Cells Cell Proliferation |
| url | http://celljournal.org/library/upload/article/af_628374225443472322342324262733245325552213-Rassouli.pdf |
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