Pollen conservation plays a crucial role in maintaining plant genetic resources, improving the efficiency of breeding programs, and facilitating germplasm conservation and exchange. However, the viability of papaya pollen gradually decreases during prolonged storage, underscoring the need for long-term preservation strategies, such as cryopreservation. This approach has gained increasing recognition because of its reliability and ability to ensure the integrity of preserved material. The maintenance of alleles crucial for crop improvement, the facilitation of germplasm exchange and institutional collaboration, and their application in cases of floral asynchrony are essential aspects of both conservation and breeding efforts. This chapter presents a case study on the cryopreservation of papaya pollen grains from accessions conserved in the Papaya Active Germplasm Bank at Embrapa Cassava and Fruits, located in Bahia, Brazil. This study details the methodological steps undertaken to establish a protocol capable of supporting the development of a cryobank for papaya pollen grains. The results obtained were robust, providing a solid foundation for the implementation of a cryobank to preserve pollen grains from different Carica papaya accessions.