Lyme Disease (LD) is a tick-borne disease caused by Borreliaburgdorferi (Bb) and transmitted in humans by Ixodesscapularis. LD can affect any organ of the body. The present study evaluated the efficacy and safety of human embryonic stem cell (hESC) therapy for the treatment of LD. Methods: Patients included in the present study had experienced symptoms of LD to varying degree of severities and intensities. The study consisted of treatment phases separated by gap phases in-between. Results: Improvement in symptoms was observed receiving hESC therapy. hESC therapy showed considerable improvement in the condition of the patients who were unable to walk straight or maintain balance while sitting and standing. These patients had regained their balance and had started to perform their regular activities with less effort after receiving hESC therapy. In addition, patients showed improvement in blurred vision, tremors, had higher energy levels, improved stamina, appetite, decreased numbness in the upper limb, decreased stiffness, regained balance, and had no slurring of speech after receiving hESC therapy. Conclusion: In conclusion, hESC therapy has shown significant improvement in patients with LD. Improvement in patients was observed without any adverse events.