| RRC ID |
85338
|
| 著者 |
Wada KI, Hosokawa K, Ito Y, Maeda M, Harada Y, Yonemitsu Y.
|
| タイトル |
Protocol for generation of transmitochondrial cybrids under pyruvate/uridine-supplemented conditions using a microfluidic device.
|
| ジャーナル |
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.
|
| 巻・号 |
6(3)
|
| ページ |
103953
|
| 公開日 |
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
|
| リソース情報 |
| ヒト・動物細胞 |
HeLa(RCB0007)
143B/TK^(-)neo^®(RCB0701) |
| 遺伝子材料 |
pTK5 (RDB01027) |
