Chen Yang^a, Jeffrey Luo^b, Marianne Polunas^c, Nikola Bosnjak^d, Sy‐Tsong Dean Chueng^b, Michelle Chadwick^e, Hatem E. Sabaawy^e, Shawn A. Chester^d, Ki‐Bum Lee^b,*, Howon Lee^a,*

^aDepartment of Mechanical and Aerospace Engineering, Rutgers University-New Brunswick 98 Brett Road, Piscataway, NJ 08854, USA
^bDepartment of Chemistry and Chemical Biology, Rutgers University-New Brunswick, 123 Bevier Rd, Piscataway, NJ 08854, USA
^cResearch Pathology Services, Rutgers University-New Brunswick, 41 Gordon Road, Suite B, Piscataway, NJ 08854, USA
^dDepartment of Mechanical Engineering, New Jersey Institute of Technology, 200 Central Ave, Newark, NJ 07102, USA
^eRutgers Cancer Institute of New Jersey, Rutgers University-New Brunswick, 195 Little Albany St, New Brunswick, NJ 08901, USA

^*To whom correspondence should be addressed.

3D cell cultures are rapidly emerging as a promising tool to model various human physiologies and pathologies by closely recapitulating key characteristics and functions of in vivo microenvironment. While high‐throughput 3D culture is readily available using multi‐well plates, assessing the internal microstructure of 3D cell cultures still remains extremely slow because of the manual, laborious, and time‐consuming histological procedures. Here, a 4D‐printed transformable tube array (TTA) using a shape‐memory polymer that enables massively parallel histological analysis of 3D cultures is presented. The interconnected TTA can be programmed to be expanded by 3.6 times of its printed dimension to match the size of a multi‐well plate, with the ability to restore its original dimension for transferring all cultures to a histology cassette in order. Being compatible with microtome sectioning, the TTA allows for parallel histology processing for the entire samples cultured in a multi‐well plate. The test result with human neural progenitor cell spheroids suggests a remarkable reduction in histology processing time by an order of magnitude. High‐throughput analysis of 3D cultures enabled by this TTA has great potential to further accelerate innovations in various 3D culture applications such as high‐throughput/content screening, drug discovery, disease modeling, and personalized medicine.