Development of cryopreservation media for the slow-freezing of cultured primordial germ cells in chicken

被引：0

作者：

Hamai, Natsuko ^{[1
,2
]}

Koide, Chihiro ^{[1
,2
]}

Tansho, Yuki ^{[1
,2
]}

Ooka, Yukino ^{[1
,2
]}

Hirano, Mayo ^{[1
,2
]}

Fatira, Effrosyni ^{[1
,2
,4
]}

Tsudzuki, Masaoki ^{[1
,2
,3
]}

Nakamura, Yoshiaki ^{[1
,2
,4
]}

机构：

[1] Hiroshima Univ, Grad Sch Integrated Sci Life, Lab Anim Breeding & Genet, Hiroshima 7398528, Japan

[2] Hiroshima Univ, Sch Appl Biol Sci, Hiroshima 7398528, Japan

[3] Hiroshima Univ, Japanese Avian Bioresource Project Res Ctr, Hiroshima 7398528, Japan

[4] Univ Palmas Gran Canaria, Unidad Asociada ULPGC CSIC, Inst Oceanog & Cambio Global, IOCAG, Parque Cientifico Tecnol Marino Taliarte, Telde 35214, Canary Islands, Spain

来源：

JOURNAL OF REPRODUCTION AND DEVELOPMENT | 2023年 / 69卷 / 02期

基金：

日本学术振兴会;

关键词：

Dimethyl sulfoxide; Primordial germ cell; Propylene glycol; Serum; Slow-freezing; trehalose; SUSTAINED-RELEASE EMULSION; LIQUID-NITROGEN; LINE CHIMERAS; COOLING RATE; STEM-CELLS; EMBRYOS; TREHALOSE; PRESERVATION; TRANSMISSION; REPLACEMENT;

D O I：

暂无

中图分类号：

S8 [畜牧、动物医学、狩猎、蚕、蜂];

学科分类号：

0905 ;

摘要：

Conservation of chicken germplasm is crucial in supporting commercial breeds for sustainable egg and meat production and preserving the genetic diversity of indigenous breeds for future breeding. Cryopreservation of chicken fertilized eggs or embryos is not feasible, owing to the large yolk-laden structure of the eggs. Primordial germ cells (PGCs), the embryonic precursors of gametes, are the best candidates for the cryobanking of chicken germplasm. Effective cryobanking of chicken PGCs requires an optimal cryopreservation protocol. Cryomedia containing dimethyl sulfoxide (DMSO) or DMSO combined with serum have been widely used for the cryopreservation of chicken PGCs. However, as cryoprotectants are yet to be optimized for chicken PGCs, the efficacy of cryomedia can be further improved. Here, we investigated the cryoprotective effects of propylene glycol (PG), an alternative to DMSO, on chicken PGCs. We found that the addition of non-permeable cryoprotectants, such as trehalose or chicken serum, to DMSO or PG cryomedia improved the recovery and survival rates of post-thawed PGCs. We further investigated the cryoprotective effects of trehalose and chicken serum and found that these additives have different cryoprotective actions. Based on these findings, we designed two different cryomedia: DTS, including 5% DMSO, 0.3 M trehalose, and 1% chicken serum, and PTS, including 7.5% PG, 0.1 M trehalose, and 5% chicken serum. Among the different PGC lines and freshly isolated PGCs, the cryomedia showed similar post-thaw recovery rates. Following transplantation, post-thawed male PGCs can colonize gonads and differentiate into functional sperm. We successfully revived the offspring of Kurokashiwa, a rare chicken breed in Japan, with cryopreserved PGCs. In conclusion, we developed two different cryomedia that achieved > 50% recovery of viable PGCs after thawing while maintaining germline competency.

引用

页码：109 / 117

页数：9