Abstract TP263: Therapeutic Potential Of Intestinal Stem Cells For Stroke Recovery: Impact Of Age Of The Donor

Nearly 50% of all stroke patients experience gastrointestinal complications that include gut microbiota dysbiosis, “leaky” gut, gut hemorrhage and gut epithelium damage. Evidence suggests that the gut signaling affects brain function and recovery from brain injury. Here we tested the hypothesis that “repairing of ischemic gut function by it’s stem cells may improve stroke outcome”. Methods: Adult and middle-aged Sprague-Dawley female and male rats were used for this study and assigned to the following groups: Control (sham), stroke with sham transplant (vehicle), or stroke with IESC transplantation. Rats were subjected to stereotaxic middle cerebral artery occlusion (MCAo) using Endothelin-1, a chemical vasoconstrictor. Primary IESCs were isolated from young or middle-aged female and male rats to prepare organoid cultures. Dispersed organoid cells were labeled with PKH67 and injected iv 4h/24h/48h to adult or middle aged hosts after stroke. Behavioral assays and saphenous blood draws were performed pre-stroke and at 2d and 4d after stroke. Trunk blood, brain tissue and a segment of small intestine were collected at termination. TTC staining was used to quantify infarct volume. Intestine samples were processed for the histomorphmetric analyses and immunohistochemistry to analyze specific markers of gut function (Lgr5+, Wnt3a, ZO-1,Ki67 and Villin). The blood-gut barrier was assessed by measuring serum levels of iFABP, LPS and Muc-2(surrogate markers of gut permeability). Results: PKH67-labelled IESC transplanted cells were observed in the basal part of the host crypt. Compared to vehicle group, MCAo-induced mortality and infarct volume were significantly reduced in middle-aged female and male rats that received IESCs from adult animals of the same sex. Sensory motor performance in the acute phase was also improved in transplanted animals as well as improved gut architecture and gut repair, and a reduction of intestinal permeability markers. However, no improvement was seen in adult animals that received senescent IESC from middle-aged animals. Conclusion: Overall, these data show that a novel stem cell population derived from the gut of young animals, but not middle-aged animals can significantly improve stroke outcomes.