Histones are a class of evolutionarily conserved nuclear proteins. Histone octamer wrapped by DNA sequence forms the nucleosome, the basic building blocks of eukaryotic chromatin. The nucleosomes keep the DNA in a condensed state, maintain the integrity of the genome, and ensure proper DNA replication, transcription, recombination and repair. Nucleosomes can regulate the biological processes of the cell through a number of distinct post-translational modifications (PTMs) and turnovers of histone variants. Although the histone H3 variant H3.3 differs from the canonical histone H3 by only a few amino acids, it could be incorporated into distinct chromatin regions by specific chaperones and exert diverse functions on the chromatin. Importantly, H3.3 is also an essential maternal factor, and plays a key role in cellular reprogramming during fertilization and somatic cell nuclear transfer. In this review, we summarize the structural properties and enrichment pattern of H3.3, and explore the specific chaperones involved in the H3.3-mediated cellular reprogramming. We hope to provide some insights on new means to improve the efficiency of somatic cell reprogramming and lay the foundation for its potential applications.