Background: Faster door to needle (DTN) times has been shown to lead to better functional outcome in patients with acute ischemic stroke. The implementation of telestroke networks has improved access to intravenous alteplase (tPA). In this study, we aim to compare DTN time between of tPA administered through telestroke in primary stroke center (PSC) spokes versus non-PSC spokes. Methods: A retrospective review of prospectively collected data on patients who received tPA through the Medical University of South Carolina telestroke program between July 2016 and March 2018. Collected data included baseline characteristics, baseline NIH stroke scale, door to CT times, door to telestroke consult time, symptom-onset to needle time, time between tPA decision and tPA administration, door to needle time, and 90-day modified Rankin scale (mRS). Wilcoxon Rank-sum (Mann-Whitney) test was used to compare median times between different groups. Results: During the study period, 399 patients received tPA through our telestroke network (314 received tPA at PSC spokes, and 85 received tPA at non-PSC spokes). PSC patients had more females (61.2%) compared to non-PSC patients (48.2%) (P=0.032). Other baseline characteristics including age and race were similar between both groups. Median NIHSS was 6 for both groups (IQR 4-9) (P=0.59). Mean times for all procedure measures were longer in non-PSC spokes; Door to CT: 8 min Vs. 5 min (P=0.029), Door to telestroke consult: 24 min Vs. 20.5 min (P=0.017), Symptom-onset to needle: 150 min Vs. 122 min (P=0.001), tPA decision to tPA administration: 20 min Vs. 10 min (<0.001), and Door to needle: 65 min Vs. 54 min (P<0.001). There was a trend towards having good functional outcome (90-day mRS 2 ) in the PSC group (79.1% and 75.7% of patients in PSC and non-PSC respectively, P=0.054) Conclusion: In our study, PSC spokes had better performance in the procedural measures for tPA administration than non-PSC spokes. Further research is needed to study the trend of these measures over time and to study the effect on the long-term functional outcome.