The preparation of self-replicating artificial cells, referred to as DNA crown cells, has been reported. Such cells could be prepared when crown cell seeds were cultured with egg white. Moreover, it was demonstrated that crown cell seeds exposed to gases could induce the emergence of living cells with agar culture. This study reports a novel in vitro method for producing living cells using gas-exposed sphingosine (Sph). Crown cell seeds consist of Sph, DNA, and adenosine and proliferated to DNA crown cells within egg white. Recently, gas-exposed Sph was used to prepare crown cell seeds instead of Sph, resulting in the in vitro emergence new living cells in seed culture. This method represents a practical and scalable alternative to previous egg-based techniques and provides a foundation for future development of synthetic cellular systems. Gas-exposed Sph was named G–Sph, and crown cell seeds prepared with G–Sph were named G–crown cell seeds. The resulting cells were named G–Sph–coli crown cells, reflecting the DNA used in the preparation of crown cell seeds.