Abstract
We recently reported isolation of viable rat amniotic fluid-derived stem (AFS) cells [1]. Here, we tested the therapeutic benefits of AFS cells in a rodent model of ischemic stroke. Adult male Sprague-Dawley rats received a 60-minute middle cerebral artery occlusion (MCAo). Thirty-five days later, animals exhibiting significant motor deficits received intravenous transplants of rat AFS cells or vehicle. At days 60–63 post-MCAo, significant recovery of motor and cognitive function was seen in stroke animals transplanted with AFS cells compared to vehicle-infused stroke animals. Infarct volume, as revealed by hematoxylin and eosin (H&E) staining, was significantly reduced, coupled with significant increments in the cell proliferation marker, Ki67, and the neuronal marker, MAP2, in the dentate gyrus (DG) [2] and the subventricular zone (SVZ) of AFS cell-transplanted stroke animals compared to vehicle-infused stroke animals. A significantly higher number of double-labeled Ki67/MAP2-positive cells and a similar trend towards increased Ki67/MAP2 double-labeling were observed in the DG and SVZ of AFS cell-transplanted stroke animals, respectively, compared to vehicle-infused stroke animals. This study reports the therapeutic potential of AFS cell transplantation in stroke animals, possibly via enhancement of endogenous repair mechanisms.
Highlights
Stroke is the fourth leading cause of death and the leading cause of disability in the United States [3]
elevated body swing test (EBST) at day 60 post-middle cerebral artery occlusion (MCAo) revealed that the swing bias was significantly decreased in the amniotic fluid-derived stem (AFS) celltransplanted stroke animals compared to the vehicle-infused stroke animals (p,0.0001) (Fig. 2A)
Rotarod testing conducted at day 60 postMCAo revealed that the AFS cell-transplanted stroke animals exhibited significantly increased time spent balancing on the rotating rod compared to the vehicle-infused stroke animals (p,0.0001) (Fig. 2B)
Summary
Stroke is the fourth leading cause of death and the leading cause of disability in the United States [3]. Extraction of AFS cells prior to delivery allows for the cells to be cultured, and in the event of childbirth-associated disorders (e.g., cerebral palsy) the stem cells can be amplified in advance and transplanted upon disease diagnosis within hours after birth This efficient amplification process may be difficult with amniotic tissue-derived cells. In the same vein of specific fate commitment, AFS cells from second trimester amniotic fluid show the capacity to differentiate into all three germ layers and expressed Oct-4, Nanog, and SSEA-4 [18], which are pluripotent embryonic stem cell markers [1,6,7,8] These findings suggest that the amniotic fluid may be an attractive source of stem cells for neurological disorders. We analyzed the effects of intravenously transplanted AFS cells in MCAo animals using cognitive and motor tests and subsequent histological analysis of the brain for determination of therapeutic benefits and mechanism of action associated with this cell therapy for stroke
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