title: Characterization of glycol chitosan grafted with low molecular weight polyethylenimine as a gene carrier for human adipose-derived mesenchymal stem cells.
abstract: Mesenchymal stem cells (MSCs) have a great capacity for self-renewal while still maintaining their multipotency, and can differentiate into a variety of cell types. The delivery of genes to a site of injury is a current and interesting field of gene therapy. In the present study, we describe a nonviral gene delivery carrier, glycol chitosan-methyl acrylate-polyethylenimine (GMP) polymer targeted towards human adipose-derived mesenchymal stem cells (AD-MSCs). Transfection efficiency, using luciferase (Luc) and a pDNA encoding enhanced green fluorescent protein (EGFP), along with cytotoxicity assays, were performed in human AD-MSCs. The results show that the transfection efficiency of the GMP polymer was similar to that of PEI25kD, and the cytotoxicity was lower. Moreover, human AD-MSCs were treated with the GMP polymer/pDNA polyplex and its cellular uptake and distribution were analyzed by flow cytometry and confocal microscopy. Furthermore, we performed endosomal escape analysis using LysoTracker Red, and found that the conjugated GMP polymer could escape from the endosome to the cytosol. Human AD-MSCs treated with the GMP polymer maintained their potential for osteogenic differentiation and phenotypic expression of human AD-MSCs based on flow cytometry analysis. The present study demonstrates that the GMP polymer can be used as a potential targeted-delivery carrier for effective gene delivery.
authors: Department of Physiology, College of Medicine, Cardiovascular and Metabolic Disease Center, Inje University, Busan 614-735, Republic of Korea.
authors: Yoonhee
authors: Y
authors: Bae
authors: Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon 305-764, Republic of Korea.
authors: Young Hwa
authors: YH
authors: Lee
authors: Cell Engineering for Origin Research Center 46-21, Susong-Dog, Jongno-Gu, Seoul 110-140, Republic of Korea.
authors: Sunray
authors: S
authors: Lee
authors: Department of Physiology, College of Medicine, Cardiovascular and Metabolic Disease Center, Inje University, Busan 614-735, Republic of Korea.
authors: Jin
authors: J
authors: Han
authors: Department of Internal Medicine, Sanggye Paik Hospital, Cardiovascular and Metabolic Disease Center, Inje University, Seoul 139-707, Republic of Korea. Electronic address: kskomd@paik.ac.kr.
authors: Kyung Soo
authors: KS
authors: Ko
authors: Department of Biochemistry, College of Natural Sciences, Chungnam National University, Daejeon 305-764, Republic of Korea. Electronic address: joonsig@cnu.ac.kr.
authors: Joon Sig
authors: JS
authors: Choi
mesh_terms: D065567:Absorption, Physiological; D000179:Acrylates / Q000737:chemistry*; D050156:Adipogenesis / Q000187:drug effects; D000273:Adipose Tissue / Q000166:cytology*; D002478:Cells, Cultured; D048271:Chitosan / Q000737:chemistry*; D003600:Cytosol; D011992:Endosomes; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins; D006801:Humans; D059630:Mesenchymal Stem Cells; D008970:Molecular Weight; D010012:Osteogenesis / Q000187:drug effects; D011094:Polyethyleneimine / Q000737:chemistry*; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry*; D014162:Transfection
chemical_list: D000179:Acrylates; D011108:Polymers; C485184:enhanced green fluorescent protein; C118638:glycol-chitosan; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine; D048271:Chitosan; C035956:methyl acrylate
keywords: Cellular uptake; Cytotoxicity; GMP polymer; Human AD-MSCs; Transfection