Preparation method and application of myocardial cell mitochondria directionally differentiated by iPSC

Highly active mitochondria were prepared by iPSC-directed differentiation of cardiomyocytes, which solved the problems of unstable donor sources and low purity, achieved effective protection against renal ischemia-reperfusion injury, reduced the risk of immune rejection, and was easy to prepare on a large scale.

CN122012383APending Publication Date: 2026-05-12THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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Patent Information

Application Number
CN202610308143.1
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2026-03-13
Publication Date
2026-05-12

AI Technical Summary

Technical Problem

Existing technologies do not provide a standardized method for preparing mitochondria of iPSC-directed differentiated cardiomyocytes, and their application in renal ischemia-reperfusion injury has not been systematically verified, resulting in unstable donor sources and low purity, making it difficult to effectively alleviate renal IRI.

Method used

Highly active mitochondria were prepared by iPSC-directed differentiation of cardiomyocytes. Cardiomyocytes with a purity ≥85% were obtained by regulating the differentiation system through the Wnt signaling pathway. Mitochondrial isolation and quality control, including transmission electron microscopy, Mito Tracker staining and cytotoxicity testing, were combined to prepare a mitochondrial formulation with a concentration of 5×10⁵ mitochondria/mL.

Benefits of technology

It significantly reduces ROS levels, restores mitochondrial membrane potential, enhances ATP synthesis capacity, reduces the proportion of apoptosis, improves renal tissue damage scores, and is easy to prepare on a large scale, avoiding the risk of immune rejection associated with cell transplantation.

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Abstract

The invention discloses a preparation method and application of IPSC directionally differentiated myocardial cell mitochondria, and relates to the technical field of mitochondria, human induced pluripotent stem cells (iPSC) are directionally differentiated into myocardial cells (iPSC-CMs), and the mitochondria is separated and extracted from the iPSC-CMs. The preparation method comprises the following steps: inducing and differentiating iPSC into iPSC-CMs by utilizing a Wnt signal channel regulation and control differentiation system, purifying by a metabolism selection method, and then separating mitochondria by a homogenate and differential centrifugation method. The mitochondria prepared by the method can be effectively ingested by renal tubular epithelial cells, and the oxidative injury of the renal tubular epithelial cells and / or the renal ischemia-reperfusion injury can be remarkably relieved by inhibiting accumulation of reactive oxygen species (ROS), improving ATP synthesis capability, recovering mitochondrial membrane potential, inhibiting apoptosis and other mechanisms.
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