Cryopreservation of bovine somatic cells using antifreeze polyamino-acid (carboxylated poly-L-lysine).
Carboxylated poly-L-lysine (CPLL) is an ampholytic polymer compound, obtained by converting 65 mol% of amino groups to carboxyl groups after synthesizing ε-poly-l-lysine aqueous solution and succinic anhydride. CPLL has cryoprotective properties similar to those of anti-freeze protein. The addition of CPLL to freezing medium has been reported to improve the post-thawing survival rate of murine cells, human induced pluripotent stem (iPS) cells, embryonic stem (ES) cells and embryos. In this study, investigating CPLL for its effectiveness as a new cryoprotective material is aimed. In experiments with bovine somatic cells, CPLL was suggested to have an equal or superior cryoprotective effect to dimethyl sulfoxide (DMSO), the conventional material for cellular frozen storage, based on the results for post-thawing cell survival and proliferation rates. CPLL was demonstrated to have another advantage; thawed cells can be cultured without removing the cryopreservation medium when CPLL is used, but not when DMSO is used. These results suggest that CPLL could be used as cryoprotective material for bovine cells. It is also expected that CPLL can be applied to embryo and oocytes storage for cattle, and similar functions for cells and embryos of other animal species.
The United Graduate School of Veterinary Science, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
Takuro
T
Fujikawa
Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065, Japan.
Takaaki
T
Ando
BioVerde.Inc., 43-1 Minamimatsunoki-cho, Higashikujo, Minami-ku, Kyoto, 601-8023, Japan.
Yuki
Y
Gen
Center for Fiber and Textile Science, Kyoto Institute of Technology (KIT), 105 Jibucho, Kyoto Fushimi-ku, Kyoto, 612-8374, Japan.
Suong-Hyu
SH
Hyon
Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065, Japan. Electronic address: k7980336@kadai.jp.
Chikara
C
Kubota
D000818:Animals; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D054885:Cumulus Cells; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005347:Fibroblasts; D011107:Polylysine / Q000031:analogs & derivatives* / Q000494:pharmacology*
D003451:Cryoprotective Agents; D011107:Polylysine; D004121:Dimethyl Sulfoxide
Bovine; Carboxylated poly-l-lysine; Cryopreservation; Dimethyl sulfoxide; Somatic cells