title: Cloning, expression, and functional characterization of in-house prepared human leukemia inhibitory factor.
abstract: 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.
  
  
  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.
  
  
  CONCLUSIONS
  Our results showed no significant differences in function between laboratory produced and commercialized hLIF.
authors.affiliation: 1. Department of Molecular Systems Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
authors.forename: Hassan
authors.initials: H
authors.lastname: Rassouli
authors.forename: Shiva
authors.initials: S
authors.lastname: Nemati
authors.forename: Siamak
authors.initials: S
authors.lastname: Rezaeiani
authors.forename: Ali
authors.initials: A
authors.lastname: Sayadmanesh
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Gharaati
authors.forename: Ghasem Hosseini
authors.initials: GH
authors.lastname: Salekdeh
authors.forename: Hossein
authors.initials: H
authors.lastname: Baharvand
authors.forename: Hamid
authors.initials: H
authors.lastname: Gourabi
keywords: Cell Proliferation; Embryonic Stem Cells; Induced Pluripotent Stem Cells; Leukemia Inhibitory Factor; Recombinant Protein