Open droplet microfluidics is an emerging technology that generates, manipulates, and analyzes droplets in open configuration systems. Droplets function as miniaturized reactors for high-throughput analysis due to their compartmentalization and parallelization, while openness enables addressing and accessing the targeted contents. The convergence of two technologies facilitates the localization and intricate manipulation of droplets using external tools, showing great potential in large-scale chemical and biological applications, particularly in cell analysis. In this review, we first introduce various methods of droplet generation and manipulation in open environments. Next, we summarize the typical applications of open droplet systems in cell culture. Then, a comprehensive overview of cell analysis is provided, including nucleic acids, proteins, metabolites, and behaviors. Finally, we present a discussion of current challenges and perspectives in this field.