Azadirachtin obtained from Azadirachta indica has garnered a wide attention as a most potent biopesticide since a long time. The limited availability of pure azadirachtin from natural trees and complex chemical synthesis protocols has led to extensive research for in vitro mass scale cultivation of A. indica suspension culture and hairy roots to meet the growing requirements of biopesticides in society. The present chapter summarizes the in vitro production strategies for azadirachtin using A. indica cell suspension and hairy root cultivations. Attempt has also been made to include the diverse bioreactor designs, impeller modifications, cultivation, and scale-up strategies for both suspension and hairy root cultures to focus on the extensive studies carried out in last two decades. The chapter also attempts to summarize the recent developments with the comprehensive analysis of genomics, transcriptomics, and proteomics studies available in the literature for A. indica.