Journal: Neuromolecular medicine

Article Title: Exosomes Secreted by the Cocultures of Normal and Oxygen-Glucose Deprived Stem Cells Improve Post-Stroke Outcome

doi: 10.1007/s12017-019-08540-y

Figure Lengend Snippet: Experimental design. Schematic diagram of the experimental design to evaluate post-stroke outcome. Rats were subjected to a right transient middle cerebral artery occlusion (tMCAO) for two hours followed by seven days reperfusion with neurological tests (mNSS, sticky-tape, and beam walking) on reperfusion days 1, 3, 5, and 7, and before ischemia. Appropriate cohorts were administered with exosomes collected under different experimental conditions intravenously via tail vein immediately after reperfusion. Rats were trained for sticky-tape and beam walk tests at least for three days prior to the induction of ischemia. Rats from few cohorts were euthanized at one-day and three-day reperfusion time points to perform TTC staining procedure

Article Snippet: Stem Cells and Collection of Exosomes Cryo-preserved human umbilical cord blood-derived mesenchymal stem cells (HUCB-MSCs) obtained from Vitro Biopharma (Golden, CO) were cultured in MSC-GRO low serum complete MSC medium and maintained according to the manufacturer’s protocol.

Techniques: Staining

Journal: Neuromolecular medicine

Article Title: Exosomes Secreted by the Cocultures of Normal and Oxygen-Glucose Deprived Stem Cells Improve Post-Stroke Outcome

doi: 10.1007/s12017-019-08540-y

Figure Lengend Snippet: Stem cells and the oxygen-glucose deprivation (OGD). a Representative bright field images of the normal and three-hour OGD-induced HUCB-MSCs in culture. b RT-PCR analysis followed by agarose gel electrophoresis. Bands represent the mRNA expression of HIF-1α. GAPDH served as a loading control. Bar graph represent the densitometry analysis of HIF-1α bands (normalized to GAPDH). Histograms and error bars indicate the mean and the SEM, respectively. n = 6. ***p < 0.001 vs control. c Detection of exosomal marker CD63 expression by immunoblot analysis. EXO-hMSCs-exosomes secreted by the hypoxic MSCs; EXO-MSCs+hMSCs – exosomes secreted by the cocultures of normal and hypoxic MSCs.

Article Snippet: Stem Cells and Collection of Exosomes Cryo-preserved human umbilical cord blood-derived mesenchymal stem cells (HUCB-MSCs) obtained from Vitro Biopharma (Golden, CO) were cultured in MSC-GRO low serum complete MSC medium and maintained according to the manufacturer’s protocol.

Techniques: Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Expressing, Marker, Western Blot

Journal: Neuromolecular medicine

Article Title: Exosomes Secreted by the Cocultures of Normal and Oxygen-Glucose Deprived Stem Cells Improve Post-Stroke Outcome

doi: 10.1007/s12017-019-08540-y

Figure Lengend Snippet: Post-stroke mortality and body weight changes. a Line graph represents the percent survival rate of rats. n=10–12. b Bar graph represents the reduction in body weight of rats at one-day reperfusion. Histograms represent the mean, and error bars indicate SEM. n=8. **p<0.01 vs. before ischemia. c Line graph represents the percent body weight gain of rats. Error bars indicate SEM. n=8–9. EXO-hMSCs - exosomes secreted by the hypoxic MSCs; EXO-MSCs+hMSCs – exosomes secreted by the cocultures of normal and hypoxic MSCs. Quantitative data of body weight gain obtained from various cohorts of rats in this study was compared with the appropriate data of vehicle-treated rats (Nalamolu et al. 2019). *p<0.05 vs. vehicle-treated; **p<0.01 vs. vehicle-treated

Article Snippet: Stem Cells and Collection of Exosomes Cryo-preserved human umbilical cord blood-derived mesenchymal stem cells (HUCB-MSCs) obtained from Vitro Biopharma (Golden, CO) were cultured in MSC-GRO low serum complete MSC medium and maintained according to the manufacturer’s protocol.

Techniques:

Journal: Neuromolecular medicine

Article Title: Exosomes Secreted by the Cocultures of Normal and Oxygen-Glucose Deprived Stem Cells Improve Post-Stroke Outcome

doi: 10.1007/s12017-019-08540-y

Figure Lengend Snippet: Assessment of the severity of post-stroke injury. Severity of the post-stroke injury at one-day reperfusion including the degree of recovery during reperfusion was assessed at regular intervals by the modified neurological severity scores (mNSS) test in rats subjected to two-hour ischemia followed by seven days reperfusion. Quantitative data of mNSS scores obtained from various cohorts of rats in this study was compared with the appropriate data of vehicle-treated rats (Nalamolu et al. 2019). Error bars indicate SEM. n=8–9. EXO-hMSCs - exosomes secreted by the hypoxic MSCs; EXO-MSCs+hMSCs – exosomes secreted by the cocultures of normal and hypoxic MSCs. *p<0.05 vs. vehicle-treated; **p<0.01 vs. vehicle-treated

Article Snippet: Stem Cells and Collection of Exosomes Cryo-preserved human umbilical cord blood-derived mesenchymal stem cells (HUCB-MSCs) obtained from Vitro Biopharma (Golden, CO) were cultured in MSC-GRO low serum complete MSC medium and maintained according to the manufacturer’s protocol.

Techniques: Modification

Journal: Neuromolecular medicine

Article Title: Exosomes Secreted by the Cocultures of Normal and Oxygen-Glucose Deprived Stem Cells Improve Post-Stroke Outcome

doi: 10.1007/s12017-019-08540-y

Figure Lengend Snippet: Effect of treatment with exosomes on infarct size and swelling. a Representative TTC stained images of rat coronal brain sections at one-day and three-day reperfusion subsequent to a two-hour focal cerebral ischemia in rats. b Bar graphs represent the quantitative data of infarct size and ipsilateral hemisphere swelling. Histograms and error bars indicate the mean and the SEM, respectively. n = 6. EXO-MSCs+hMSCs – exosomes secreted by the cocultures of normal and hypoxic MSCs. Quantitative data of infarct size and swelling obtained from EXO-MSCs+hMSCs treated rats in this study was compared with the appropriate data of untreated, ischemia-induced rats that we reported earlier (Chelluboina et al. 2015; Nalamolu et al. 2018b). **p<0.01 vs. untreated ischemia-induced rats euthanized at one-day reperfusion; ###p<0.001 vs. untreated ischemia-induced rats euthanized at three-day reperfusion

Article Snippet: Stem Cells and Collection of Exosomes Cryo-preserved human umbilical cord blood-derived mesenchymal stem cells (HUCB-MSCs) obtained from Vitro Biopharma (Golden, CO) were cultured in MSC-GRO low serum complete MSC medium and maintained according to the manufacturer’s protocol.

Techniques: Staining

Journal: Neuromolecular medicine

Article Title: Exosomes Secreted by the Cocultures of Normal and Oxygen-Glucose Deprived Stem Cells Improve Post-Stroke Outcome

doi: 10.1007/s12017-019-08540-y

Figure Lengend Snippet: Assessment of the post-stroke somatosensory dysfunction and deficits in coordination and integration of motor movement. Somatosensory dysfunction (a), and deficits in coordination and integration of motor movement (b) at one-day reperfusion including the degree of recovery during reperfusion were assessed at regular intervals by the modified adhesive removal (sticky-tape) test, and the beam walking test, respectively, in rats subjected to two-hour ischemia followed by seven days reperfusion. Quantitative data of sticky-tape ratio and beam walking scores obtained from various cohorts of rats in this study was compared with the appropriate data of vehicle-treated rats (Nalamolu et al. 2019). Error bars indicate SEM. n=7–9. EXO-hMSCs - exosomes secreted by the hypoxic MSCs; EXO-MSCs+hMSCs – exosomes secreted by the cocultures of normal and hypoxic MSCs. *p<0.05 vs. vehicle-treated; **p<0.01 vs. vehicle-treated

Article Snippet: Stem Cells and Collection of Exosomes Cryo-preserved human umbilical cord blood-derived mesenchymal stem cells (HUCB-MSCs) obtained from Vitro Biopharma (Golden, CO) were cultured in MSC-GRO low serum complete MSC medium and maintained according to the manufacturer’s protocol.

Techniques: Modification