Background: Interstitial cystitis/bladder pain syndrome (IC/BPS) is a chronic and diagnostically challenging condition with limited treatment options and poor prognosis. Platelet-rich plasma (PRP) is a potential treatment for IC/BPS. Nonetheless, conventional preparation methods are inefficient, necessitating an improved approach. Objective: To design an effective and safe protocol for collecting PRP for intravesical injections for the treatment of IC/BPS. Methods: We evaluated the feasibility of the PRP collection protocol from 17 patients. After venipuncture, blood was processed using a blood cell separator containing saline and anticoagulant. One milliliter of PRP was retained for platelet concentration analysis. Each patient received six PRP injections, and their clinical data were taken before and after the procedure to assess efficacy. Results: The mean platelet enrichment coefficient was 5.11 ± 1.27, the mean flow rate was 33.19 ± 6.77 mL/min, the mean collection volume was 125.13 ± 17.49 mL, and the mean collection time was 73.69 ± 10.17 min. Eleven patients required deep venous catheterization because superficial venipuncture failed. No adverse reactions were observed during collection, and all other blood components were retained. Three patients completed six injections, four received five injections, three received four injections, one received three injections, two received two injections, and four received one injection. After treatment, 12 of 17 patients showed symptom improvement on the Global Response Assessment scale (≥5), and all demonstrated statistically significant improvements in terms of O’Leary-Sant and Visual Analog Scale scores against baseline. One patient, however, developed gross hematuria after the fourth injection. Conclusions: The proposed PRP collection protocol is relatively safe and effective. Preliminary data suggest that intravesical PRP injection is a promising treatment for IC/BPS.