ISSCR 2019: 26-29 JUNE

LOS ANGELES

(W-3225)

STRUCTURAL AND MOLECULAR-BASED DIFFERENTIAL FEATURES BY LONG TERM-INDUCED MESENCHYMAL STEM CELLS IN A RAT MODEL OF PARKINSONS DISEASE

Song, Byeong-Wook - Department for Medical Sciences, Catholic Kwandong University, Incheon, Korea

Kim, Il-Kwon - Institute for Bio-Medical Convergence, Catholic Kwandong University, Incheon, Korea

Lee, Jiyun - Institute for Bio-Medical Convergence, Catholic Kwandong University, Incheon, Korea

Kang, Misun - Institute for Bio-Medical Convergence, Catholic Kwandong University, Incheon, Korea

Park, Jun-Hee - Institute for Bio-Medical Convergence, Catholic Kwandong University, Incheon, Korea

Choi, Jung-Won - Institute for Bio-Medical Convergence, Catholic Kwandong University, Incheon, Korea

Kim, Sang Woo - Institute for Bio-Medical Convergence, Catholic Kwandong University, Incheon, Korea

Lim, Soyeon - Institute for Bio-Medical Convergence, Catholic Kwandong University, Incheon, Korea

Lee, Seahyoung - Institute for Bio-Medical Convergence, Catholic Kwandong University, Incheon, Korea

Hwang, Ki-chul - Institute for Bio-Medical Convergence, Catholic Kwandong University, Incheon, Korea

Adult stem cell is a therapeutic source for restoring neuronal function in brain disease. To improve Parkinson’s disease (PD) belonging to neurodegenerative disease, stem cell therapy is known to functionally enhance nigrostriatal dopaminergic neurons. Despite these functional recoveries, the long-term state of stem cells in vivo and their relationship with surrounding cells have not been clearly studied. In previous study, we examined a three dimensional structure of intact brain tissue after bone marrow mesenchymal stem cell (MSC) treatment at short-term induction in PD. In this study, we investigated in vivo Proof-of-concept and their microenvironment of MSCs 1 year after transplantation into 6-hydroxydopamine hydrobromide-induced rat PD model at the medial forebrain. One year after surgery, amphetamine-induced rotation test was performed to evaluate the degree of the dopaminergic lesion. For transplantation, MSCs were injected into the femoral vein. One day after the final behavior test, the rats were anaesthetized, and perfused with a mixture of 4% paraformaldehyde, 4% acrylamaide, 0.05% bisacrylamide, 0.25 VA044 in PBS. Isolated brains were incubated in 4°C for 2 days, and then temperature was increased to 37°C to induce polymerization for 2.5 hours. Hydrogel-embedded brains were plated in 10~60V organ-electrophoresis system, circulating 200 mM sodium borate buffer with 4% SDS. For three dimensional interface of graft-host axis, it was visualized by immunofluorescence to MSC-positive cells (PKH26-labelled), polysialylated neural cell adhesion molecule, and/or neuronspecific class III beta-tubulin or NeuN marker. At this site, we analyzed RNA-Sequencing to confirm the change of gene expression between PD and PD transplanted with MSC group. Using Excel-based Differentially Expressed Gene Analysis tool, we analyzed the categories related to cell migration and neurogenesis between the two groups. Furthermore, GO and Pathway and Network Analysis was mainly performed on the specifically changed genes. Taken together, our results demonstrate distinct pathophysiological and molecular patterns of in vivo responses between PD and PD transplanted with MSCs suggest structural and molecular-based differential features as candidate tool for the evaluation of stem cell during long-term therapy.

Funding Source: This study was funded by NRF-2015R1C1A1A02037693 and NRF-2015M3A9E6029519.​