Automated Optical Tweezers Manipulation to Transfer Mitochondria from Fetal to Adult MSCs to Improve Antiaging Gene Expressions

Abstract

Mitochondrial dysfunction is considered to be an important factor that leads to aging and premature aging diseases. Transferring mitochondria to cells is an emerging and promising technique for the therapy of mitochondrial deoxyribonucleic acid (mtDNA)‐related diseases. This paper presents a unique method of controlling the quality and quantity of mitochondria transferred to single cells using an automated optical tweezer‐based micromanipulation system. The proposed method can automatically, accurately, and efficiently collect and transport healthy mitochondria to cells, and the recipient cells then take up the mitochondria through endocytosis. The results of the study reveal the possibility of using mitochondria from fetal mesenchymal stem cells (fMSCs) as a potential source to reverse the aging‐related phenotype and improve metabolic activities in adult mesenchymal stem cells (aMSCs). The results of the quantitative polymerase chain reaction analysis show that the transfer of isolated mitochondria from fMSCs to a single aMSC can significantly increase the antiaging and metabolic gene expression in the aMSC. The proposed mitochondrial transfer method can greatly promote precision medicine for cell therapy of mtDNA‐related diseases.

High-efficiency quantitative control of mitochondrial transfer based on droplet microfluidics and its application on muscle regeneration

“…The number of mitochondria transferred into recipient cells can be controlled by adjusting the concentration of the isolated mitochondria suspension used. Compared with the traditional coculture method ( 16 , 24 ) and the recently proposed OT-based method ( 14 ), the method proposed in the present study can achieve quantitative control of mitochondrial transfer into recipient cells at the single-cell level while maintaining a high mitochondria transfer efficiency of 75% and a high throughput, generating 2 × 10 6 mitochondria-transferred cells in 2.5 hours (0.5 hours for generating droplets containing the cells and isolated mitochondria and 2 hours for the mitochondrial transfer process in droplets). The previous work showed that the number of mitochondria transferred into recipient cells increased with the time used for the transfer process.…”

“…Despite being a rather simple process, the success of the coculture method is dependent on many uncontrollable factors, which might determine the unsatisfactory cell metabolism recovery rate of recipient cells in a previous work (~0.2%) ( 16 ). An automated optical tweezers (OT)–based manipulation system was used for qualitative and quantitative mitochondrial transfer ( 14 ) to reduce the influence from uncontrollable factors. The OT-based system could precisely pick up the healthy mitochondria and transport them to the target recipient cell.…”

“…Apart from energy production for cells, mitochondria can influence cell proliferation, aging, apoptosis, innate immunity, calcium homeostasis, and even stem cell differentiation potential (12)(13)(14). Mutations of mtDNA impair the functions of cells and tissues.…”

Advanced tools and methods for single-cell surgery

“…OTs have been effectively used to investigate numerous biological applications in recent years 21 , 59 , 60 . As demonstrated in Fig.…”

A novel methodology for detecting variations in RBCs surface antigens by cell-tearing using optical tweezers

“…Subsequently, the OT is now being used in the investigation of an increasing number of biochemical and biophysical processes [23][24][25][26][27][28][29][30][31][32][33]. After decades of developments, convenient commercial OT machines and automatic manipulation systems with microfluidic chips [34][35][36][37] are becoming available and popular. Many easy-to-use OT products have been promoted.…”