Abstract Details

Title A Therapeutic Approach Using Peripheral Blood Mononuclear Cells Preconditioned by Oxygen-glucose Deprivation in Ischemic Rats

Topic Cerebrovascular Disease and Interventional Neurology

Presentation(s) P1 - Poster Session 1 (12:00 PM-1:00 PM)

Poster/Presentation
Number 4-003

Objective To determine the effects of administration of peripheral blood mononuclear cells (PBMCs) preconditioned by oxygen-glucose deprivation (OGD-PBMCs) for ischemic stroke.

Background Cell therapies that invoke pleiotropic mechanisms facilitating functional recovery in stroke patients. Based on previous experiments using microglia, we hypothesized that optimal OGD condition must be suitable for switching PBMCs to the protective polarisation state in microglia.

Design/Methods We prepared primary PBMCs from wild-type rats and human by the Ficoll centrifugation. We compared levels of growth factor, vascular endothelial growth factor (VEGF), and cytokines in PBMCs-conditioned media under normoxic and OGD conditions. Then, we performed confocal microscopic analyses to assess the expression level of remodeling factors, angiogenesis, and axonal outgrowth by administration of OGD-PBMCs after ischemic stroke using the suture technique in rats. Finally, we examined the therapeutic benefits of intra-arterially administered OGD-PBMCs after focal cerebral ischemia.

Results We confirmed marked secretion of remodeling factors in vitro. First, while VEGF levels were found in the conditioned media of OGD-PBMCs, the same was not valid for PBMCs under the normoxic condition (P=0.03). Expression of the anti-inflammatory cytokine, transforming growth factor-β (TGF-β), was twice higher after OGD compared with a normoxic condition (P=0.04), and the ratio of TGF-β per tumor necrosis factor-α, which shows the polarization of protective anti-inflammatory and toxic pro-inflammatory monocytes/macrophages and microglia, was five times higher after OGD, compared with a normoxic condition (P=0.04). Thus, we found that intra-arterial administration of OGD-PBMCs caused increased expression of VEGF and TGF-β around the injured brain parenchyma. This treatment promoted angiogenesis as well as axonal outgrowth in the ischemic penumbra. Finally, we demonstrated that administration of OGD-PBMCs at 7 days after ischemia prompted functional recovery at 28 days after focal cerebral ischemia compared with control therapies.

Conclusions Intravascular administration of PBMCs preconditioned by OGD might be a novel therapeutic strategy against ischemic stroke.

Authors/Disclosures
Masato Kanazawa, MD, FAAN (Niigata University of Health and Welfare) PRESENTER	Dr. Kanazawa has nothing to disclose.
	No disclosure on file
Tetsuya Takahashi, PhD (Dept. of Neurology, Niigata University BRI)	No disclosure on file
	No disclosure on file
	No disclosure on file
Tetsuya Takahashi, PhD (Dept. of Neurology, Niigata University BRI)	No disclosure on file
Osamu Onodera, MD	Dr. Onodera has nothing to disclose.
	No disclosure on file
Takayoshi Shimohata, MD, FAAN (Department of Neurology, Gifu University)	Dr. Shimohata has nothing to disclose.