Nanofiber membranes (NFMs), which have an extracellular matrix (ECM)‐mimicking structure and unique physical properties, have garnered great attention as biomimetic materials for developing physiologically relevant in vitro organ/tissue models. Recent progress in NFM fabrication techniques immensely contributed to the development of NFM‐based cell culture platforms for constructing physiological organ/tissue models. However, despite the significance of the NFM fabrication technique, an in‐depth discussion of the fabrication technique and its future aspect was insufficient. This review provides an overview of the current state‐of‐the‐art of NFM fabrication techniques from electrospinning techniques to post‐processing techniques for the fabrication of various types of NFM‐based cell culture platforms. Moreover, the advantages of the NFM‐based culture platforms in the construction of organ/tissue models are discussed especially for tissue barrier models, spheroids/organoids, and biomimetic organ/tissue constructs. Finally, the review concludes with perspectives on challenges and future directions for fabrication and utilization of NFMs.This article is protected by copyright. All rights reserved