Protective effect of both human TH and GDNF cDNA-engineered SH-SY5Y cells on primary dopaminergic neurons / 中华老年医学杂志

ABSTRACT

Objective To construct genetically engineered cells that secrete human TH and GDNF at the same time by stable co-transfection pcDNA3.0/hTH and pcDNA3.1/hGDNF in SH-SY5Y cells and study their effects on the gene therapy of Parkinson′s disease (PD). Methods pcDNA3.0/hTH and pcDNA3.1/hGDNF were constructed by inserting human TH and GDNF cDNA into pcDNA3.0 and pcDNA3.1, respectively. SH-SY5Y cells were transfected with pcDNA3.0/hTH and pcDNA3.1/hGDNF and the positive cell clones of human TH and GDNF cDNA engineered cells could be identified by RT-PCR. The engineered cells were co-cultured with primary dopaminergic neurons in mouse. The number and growth condition of primary dopaminergic neurons were examined by Immunocytochemistry. Results The number of primary dopaminergic neurons protected by double-gene engineered cells increased at least by 2.8 times in comparison with the control cells(P