Open AccessLocalized SDF‐1alpha gene release mediated by collagen substrate induces CD117 + stem cells homing
Author(s) -
Wang Weiwei,
Li Wenzhong,
Ong LeeLee,
Furlani Dario,
Kaminski Alexander,
Liebold Andreas,
Lützow Karola,
Lendlein Andreas,
Wang Jun,
Li RenKe,
Steinhoff Gustav,
Ma Nan
Publication year - 2010
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/j.1582-4934.2008.00624.x
Subject(s) - stem cell , transfection , microbiology and biotechnology , chemistry , stromal cell , homing (biology) , biology , biochemistry , cancer research , gene , ecology
Stromal cell‐derived factor‐1α (SDF‐1α) mediated mobilization and homing of stem cells showed promising potential in stem cell based tissue engineering and regenerative medicine. However local and sustained release of SDF‐1α is indispensable for stem cell mediated regenerative process due to its short half‐life under inflammatory conditions. In this study, a gene activated collagen substrate (GAC) was formed via assembly of plasmid encoding SDF‐1α into a collagen substrate to create a microenvironment favoring stem cell homing. Local release of SDF‐1α from the transfected cells on GAC and its effect on CD117 + stem cell homing were investigated. Non‐viral poly‐ethyleneimine (25kDa PEI)/DNA complexes were mixed with rat tail collagen solution to form the GAC. Optimization of GAC was carried out based on collagen effects on the PEI/DNA complexes, viability and luciferase expression of COS7 cells on GAC. CD117 + stem cells homing in response to SDF‐1α local expression from transfected cells on GAC were investigated in a flow chamber in vitro and in a mouse hind limb model in vivo . The gene expression, migration of CD117 + stem cells and the induced inflammation were investigated with immunostaining, reverse transcription polymerase chain reaction (RT‐PCR) and H&E staining. The optimized parameters for GAC were DNA dosage 10 μg/cm 2 , molar ratio of PEI nitrogen in primary amine to DNA phosphate (N/P ratio) 4 and mass ratio of collagen to DNA (C/D ratio) 1.0. It kept cell viability above 75% and transfection efficiency around 5.8 × 10 5 RLU/mg protein. GAC allowed the sustained gene release up to 60 days. GAC mediated SDF‐1α gene release induced migration and homing of CD117 + stem cells in vitro and in vivo significantly, and the inflammation of GAC reduced significantly two weeks after transplantation. GAC is a promising stem cell based therapeutic strategy for regenerative medicine.