Abstract: Cancer is a group of heterogeneous diseases caused by diverse genomic alterations in oncogenes and tumor suppressor genes. Despite recent advances in high-throughput sequencing technologies and the development of targeted cancer therapy, novel cancer drug development is limited due to the high attrition rate from clinical studies. Tumor cell line culture technologies have emerged as a strong impetus to the development of tumor biology research. However, cell line tumor models present limited predictive value and unstable properties in the clinical translational research of new cancer therapies. Conditional reprogrammed cells (CRCs) allow the establishment of normal and tumor epithelial cell cultures from specific tissue types of patients, thus accurately reflecting the heterogeneity of original tumor cells and the diversity of genetic information for preclinical pharmacology of personalized screening assessment and clinical routine targeted therapy after drug-resistance. This provides an effective research resource for new drug discovery. This review summarizes the progress of CRC research, including its application in tumor neoadjuvant therapy, challenges, limitations, and its application prospects in precision cancer medicine.