| RRC ID |
85338
|
| Author |
Wada KI, Hosokawa K, Ito Y, Maeda M, Harada Y, Yonemitsu Y.
|
| Title |
Protocol for generation of transmitochondrial cybrids under pyruvate/uridine-supplemented conditions using a microfluidic device.
|
| Journal |
STAR Protoc
|
| Abstract |
Under conventional approaches, selective culture with a pyruvate/uridine (PU)-free medium is essential for generating transmitochondrial cybrids. Here, we present a protocol for generating transmitochondrial cybrids using a microfluidic device, which works even under PU-supplemented conditions. We describe steps for preparation of the microfluidic device, partial cell fusion (mitochondrial transfer), and harvest of transmitochondrial cybrids. We then detail procedures for confirmation of mtDNA repopulation in cybrids and mtDNA typing by restriction fragment length polymorphism (RFLP) analysis. For complete details on the use and execution of this protocol, please refer to Wada et al.1.
|
| Volume |
6(3)
|
| Pages |
103953
|
| Published |
2025-7-17
|
| DOI |
10.1016/j.xpro.2025.103953
|
| PII |
S2666-1667(25)00359-4
|
| PMID |
40682780
|
| PMC |
PMC12296426
|
| MeSH |
DNA, Mitochondrial / genetics
Humans
Lab-On-A-Chip Devices*
Mitochondria* / genetics
Mitochondria* / metabolism
Polymorphism, Restriction Fragment Length
Pyruvic Acid* / metabolism
Uridine* / metabolism
|
| Resource |
| Human and Animal Cells |
HeLa(RCB0007)
143B/TK^(-)neo^®(RCB0701) |
| DNA material |
pTK5 (RDB01027) |
