In-vitro androgenesis offers a distinctive approach for generating homozygous doubled-haploid plants. Utilizing haploid plant breeding speeds up the process of obtaining doubled haploid plants, which plays a crucial role in rice breeding. This research aims to enhance the production of doubled haploids through In-vitro anther culture in rice and to select doubled haploid plants with desirable traits.The research finds that a 9-day cold treatment positively influences callus formation when using modified N6 media, whereas MS media is applied for plant regeneration. Six plant regeneration media (PRM1, PRM2, PRM3, PRM4, PRM5, and PRM6) were tested, with PRM5 (BAP 0.5 mg/L, NAA 0.25 mg/L) and PRM6 (BAP 1.5 mg/L, NAA 1.0 mg/L, kinetin 1.0 mg/L) with successful plant regeneration. PRM6 showed the highest regeneration rate with 12 plants, while PRM5 resulted in 8 plants. Other media (PRM1–PRM4) led only to callus blackening, with no plant regeneration. A total of 20 albino plants were developed, demonstrating that higher growth regulator concentrations may increase callus regrowth under these conditions. A major challenge observed in this work was the complete regeneration of albino plants, indicating impaired chloroplast development. The study contributes to rice androgenesis research by identifying effective cold pre-treatment durations, evaluating the hormonal combinations for callus proliferation and regeneration, and highlighting critical bottlenecks such as albinism that must be addressed in future protocol optimization.