Preparation method and application of conditionally immortalized human myocardial fibroblast model

By constructing a lentiviral vector carrying the SV40LT-tsA58 gene regulated by the Tet-on system and combining it with temperature-sensitive mutations, a conditionally immortalized human cardiomyocyte fibroblast model was achieved. This solved the problems of time-consuming and labor-intensive preparation, large heterogeneity, ethical issues, and functional differences in existing technologies, and provided an efficient and reliable drug screening tool.

CN121950706APending Publication Date: 2026-05-01SHENZHEN LONGGANG DISTRICT PEOPLES HOSPITAL
0 Cites 0 Cited by

Patent Information

Application Number
CN202511967531.2
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-12-24
Publication Date
2026-05-01

AI Technical Summary

Technical Problem

Existing cardiac fibroblast cell models suffer from problems such as being time-consuming and labor-intensive in preparation and use, high cost, large heterogeneity, ethical issues, and difficulty in accurately simulating human disease characteristics. Furthermore, traditional immortalization methods result in significant differences in cell function, making it impossible to truly simulate physiological and pathological states.

Method used

Human cardiomyocytes were transduced using a lentiviral vector carrying the SV40LT-tsA58 gene, which is regulated by the Tet-on system. Conditional immortalization was achieved by driving the expression of the tetR-KRAB fusion protein through the cardiomyocyte-specific promoter COL1A2. The model was then combined with the temperature-sensitive mutated SV40LT-tsA58 gene for controlled proliferation, ensuring the safety and reversibility of the model.

Benefits of technology

It provides an efficient and reliable conditionally immortalized human cardiomyocyte model that can simulate physiological and pathological states with high fidelity, achieve high-throughput drug screening, reduce costs, avoid ethical issues, and ensure system safety and reliability through a dual control switch.

✦ Generated by Eureka AI based on patent content.
Patent Text Reader

Abstract

The invention relates to the field of cell engineering, and particularly discloses a preparation method and application of a conditionally immortalized human myocardial fibroblast model. According to the method, a Tet-on system driven by a cardiac fibroblast specific promoter is constructed to regulate and control a lentiviral vector of an SV40LT-tsA58 temperature-sensitive mutant gene, and after primary human cardiac fibroblasts are transduced, conditional immortalization of the cells is realized in the presence of an inducer doxycycline. The model can be stably amplified under an induction condition, the immortalized gene expression is closed after an inducer is removed, the primary physiological state of cells is recovered, and the high-fidelity response capability to fibrosis promoting and anti-fibrosis drugs is reserved. According to the method, the in-vitro proliferation limit of primary cells is broken through, the problems of species difference and ethics are avoided, and a humanized, high-fidelity and high-controllability standardized tool is provided for myocardial fibrosis pathological mechanism research and drug screening.
Need to check novelty before this filing date? Find Prior Art