With the advancement of human spaceflight into deep space, space life science has become a pivotal field for studying the effects of the cosmic environment on living organisms. Space-specific environmental factors—such as microgravity and galactic cosmic radiation—exert profound impacts on life across multiple levels, from molecular to systemic. However, limitations of traditional research models and in situ detection technologies have long hindered in-depth mechanistic understanding. In recent years, the integration of microphysiological systems (including organoids and organ-on-chips) with advanced in-orbit detection technologies is driving a fundamental transformation in the research paradigm of space life science. This article systematically reviews the unique advantages of microphysiological systems in mimicking the three-dimensional structure and physiological functions of human organs, and summarizes their successful applications onboard the International Space Station and other platforms, covering research progress and key findings in brain, bone, immune tissue, and other tissue models. Furthermore, it provides a detailed evaluation of recent advances in in situ detection technologies such as high-content fluorescence imaging, light-sheet microscopy, Raman spectroscopy, and nanopore sequencing, which together enable real-time, dynamic, and multi-modal monitoring of biological processes in space. The article also analyzes major current challenges in the field, including limited technological integration, insufficient long-term culture systems, and difficulties in multi-modal data fusion. Finally, it outlines future directions for building intelligent and integrated space experimental platforms, emphasizing that the deep integration of multi-modal sensing, artificial intelligence, and automation will advance space life science into a new era of multi-scale, systematic, and precise analysis.