New methods have been developed which streamline microbial seed train processes and offer the potential to decrease costs and thereby provide a competitive advantage.  One of the methods that have been previously used in microbial fermentation processes for biopharmaceutical production at pilot scale, with success, is the use of frozen bags of inoculum rather than shake flasks and seed fermenters.  This approach has been scaled down for 20 L microbial fermentation development and has an obvious advantage in reducing cycle time as an inoculum bag, depending on size, may take a few hours or less to thaw and be ready to use.  A typical seed train for any given microbial fermentation process can take from half a day to five days or greater to develop depending on the organism and other growth conditions.  Another advantage of this method is the ability to create high density seeds that can be adjusted to influence the length of fermentation time.  Aside from time advantages, this technique conserves cell bank resources, reduces effort needed to produce seed trains, reduces fermentation process variability due to seeding inoculum differences, and ultimately saves money by reducing resources needed to produce seed trains for discrete fermentation batches