title: Lentivirus vector-mediated gene transduction of CNGRC peptide in rat adipose stem cells.
abstract: The aim of the present study was to investigate the feasibility of lentiviral‑mediated Cys‑Asn‑Gly‑Arg‑Cys (CNGRC) peptide gene transduction in adipose stem cells. Adipose stem cells were prepared using enzymatic digestion and repeated adherence methods and identified in culture by immunofluorescence staining of surface markers. The pluripotency of the cultured adipose stem cells was confirmed by their induced differentiation into bone and fat cells. Following polymerase chain reaction amplification, the gene sequence for the CNGRC peptide was cloned into a lentiviral vector, which was then co‑transfected into 293T cells with packaging plasmids Helper 1.0 and Helper 2.0. The lentiviruses carrying the CNGRC peptide gene were then harvested and used to transfect adipose stem cells. Following transduction, expression of CNGRC in adipose stem cells was detected using western blot analysis. Adipose stem cells in culture were successfully induced to differentiate into adipocytes and osteoblasts and the lentiviral vector containing CNGRC‑3Flag‑EGFP was successfully constructed. Following transduction, western blot analysis and immunofluorescence staining demonstrated expression of the CNGRC protein in adipose stem cells. This suggested that adipose stem cell lines expressing the CNGRC peptide were successfully established.
authors.affiliation: Department of Oncology, First Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
authors.forename: Du
authors.initials: D
authors.lastname: Meng
authors.affiliation: Department of Oncology, First Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
authors.forename: Rui
authors.initials: R
authors.lastname: Liu
authors.affiliation: Department of Oncology, First Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
authors.forename: Li
authors.initials: L
authors.lastname: Pei
authors.affiliation: Department of Oncology, First Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
authors.forename: Lei
authors.initials: L
authors.lastname: Hou
authors.affiliation: Department of Oncology, First Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Ning
authors.affiliation: Department of Oncology, First Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Yu
authors.affiliation: Department of Oncology, First Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
authors.forename: Lu
authors.initials: L
authors.lastname: Feng
authors.affiliation: Department of Oncology, First Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
authors.forename: Xinhan
authors.initials: X
authors.lastname: Zhao
mesh_terms: D017667:Adipocytes / Q000166:cytology; D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D000954:Antigens, Surface / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D016130:Immunophenotyping; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D010011:Osteocytes / Q000166:cytology; D010456:Peptides, Cyclic / Q000235:genetics*; D010641:Phenotype; D051381:Rats; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D014161:Transduction, Genetic
chemical_list: D000954:Antigens, Surface; D010456:Peptides, Cyclic