Effect of human amniotic mesenchymal stem cell transplantation on motor behaviors in hSOD1-G93A mouse models of amyotrophic lateral sclerosis

Abstract

Objective To explore the therapeutic effect of intravenous transplantation of human amniotic mesenchymal stem cells (hAMSCs) on protection of motor behaviors in hSOD1-G93A mouse models of amyotrophic lateral sclerosis (ALS). Methods Amnion membranes were obtained from placentas delivered by healthy mother donors. The hAMSCs were gradually isolated and purified from amnion membranes using tissue culture method. Immunophenotypes of the isolated hAMSCs were analyzed using fluorescence activated cell sorter (FACS). Transgenic mice harboring a high copy number of hSOD1-G93A (B6SJL-TgN [SOD1-G93A] 1Gur) transgene were used in this study. Hemizygous transgenic progenies were maintained by mating the transgenic males with F1 hybrid wild-type (WT) females. The progenies were genotyped by polymerase chain reaction (PCR) using genomic DNA isolated from mouse tail after birth. The study included hSOD1-G93A mice transplanted with hAMSCs, PBS-injected transgenic mice, and normal WT mice (n=12). The hAMSCs were administered intravenously in jugular vein of the mice under anesthesia. The cells (1×106) in 200 μL PBS were delivered over 10 min. Animals received cells or PBS at 12, 14, and 16 weeks old, respectively. The disease onset and progression of ALS mice models were monitored using rotarod performance test, PaGE test, and CatWalk gait analysis since 8 weeks old every week. Results (1) The immunophenotype of the isolated hAMSCs was conformed using FACS. These cells were positive for CD29, CD44, CD73, CD90, and CD166, but negative for CD14, CD34, CD45, CD123, and human leukocyte (site) DR antigen. Interestingly, stage specific embryo surface antigen 4 and octuber binding transcription factor 4 were detected in hAMSCs. (2) ALS mice in the hAMSCs transplantation group had significantly improved motor functions than those in the PBS treatment group: motor performance on the rotarod test (from 14 to 18 weeks old), PaGE test (from 15 to 18 weeks old) and CatWalk gait analysis (from 15 to 19 weeks old) in hAMSCs-injected ALS mice was significantly improved as compared with that in the PBS treatment group (P<0.05). Conclusions The multiple transplantation of hAMSCs by intravenous delivery can bring amelioration of the disease phenotype, as evidenced by improved motor function in hSOD1-G93A mouse models. The hAMSCs transplantation can be considered as a promising cellular treatment for ALS. Key words: Amyotrophic lateral sclerosis; Human amniotic mesenchymal stem cell; Stem cell transplantation