The production of haploid/doubled haploid plants allows one to speed up breeding programs, improve selection efficiency, detect linkage and gene interactions, estimate genetic variance and the number of genes for quantitative characteristics, produce genetic translocations, substitutions and chromosome addition lines and facilitate genetic transformation. In breeding programs, besides doubled haploids production, the microspore culture can be used to produce and maintain malesterile plants via microspore embryogenesis, possibility to restore male-fertility via in vitro microspore maturation, to overcome self-incompatibility and to induce and select for mutants. In genetic engineering, Male Germ Line Transformation (MAGELITR) in which DNA is transferred into microspores and cultured in vitro to form mature pollen and then using the transformed pollen for pollination in vivo, is very efficient method. In basic studies, the microspore culture can be used to investigate pollination, pollen development, embryogenesis, totipotency, cell cycle, phase change and the role of stress in development. In this paper, various aspects of in vitro microspore culture are discussed.