Pluripotency of cryopreserved blastomeres of the goldfish

Abstract

To examine the pluripotency of cryopreserved blastomeres, we transplanted them into blastula. Donor blastomeres were prepared from blastula of goldfish (Carassius auratus) and cryopreserved in liquid nitrogen for two months. Fifty-five percent and 44% of blastomeres survived after thawing. Cryopreserved blastomeres were transplanted to the blastula of triploid crucian carp (C. a. longsdorfii), which reproduces gynogenetically in nature. At four days after the operation, resultant chimeric embryos transplanted with cryopreserved blastomeres showed a survival rate (41.6%) lower than that of embryos transplanted with unfrozen blastomeres (57.1%). Transplanted blastomeres were histologically identified in various organs derived from all three germ layers. A primordial germ cell differentiated from a cryopreserved blastomere was detected in one of the 32 chimeric fish examined. These results suggest blastomeres that survive after cryopreservation retain their pluripotency and are able to differentiate into both somatic and germ cell lines.

Developmental biotechnology for aquaculture, with special reference to surrogate production in teleost fishes

“…The cryopreservation of dissociated blastomeres from the blastula was also carried out in several species (Nilsson and Cloud, 1993;Kusuda et al, 2004). The survival of cryopreserved blastomeres was about 50% after thawing (Kusuda et al, 2002(Kusuda et al, , 2004.…”

“…The survival of cryopreserved blastomeres was about 50% after thawing (Kusuda et al, 2002(Kusuda et al, , 2004. In goldfish, when blastomeres were transplanted into the host embryo after thawing, a PGC derived from donor blastomeres was detected in the resultant embryo (3%, 1/32) (Kusuda et al, 2004). This low frequency of germ-line chimerism was due to the relatively low number of PGCs per embryo at the blastula stage.…”

“…This low frequency of germ-line chimerism was due to the relatively low number of PGCs per embryo at the blastula stage. The method of blastomere cryopreservation by Kusuda et al (2004) may be available for isolated PGCs after the establishment of a mass isolation technique for PGCs.…”

“…In teleost species, pedigree preservation and sperm cryopreservation have been carried out in a relatively large number of species Jamieson, 1991, Kusuda, 2005). However, genetic diversity decreases during pedigree preservation, and maternal genes are not preserved in sperm cryopreservation (Kusuda et al, 2004). Cryopreservation of PGCs and the recovery of species through surrogate parents of related species are useful for the preservation of genetic diversity, including cytoplasmic genes .…”

Technical note: Viability and motility of vitrified/thawed primordial germ cell isolated from common carp (Cyprinus carpio) somite embryos1

“…In these studies, mixed EG and DMSO were used as cryoprotectants. In previous studies, EG and DMSO have been the cryoprotectants of choice for preserving the embryos of a range of species (Kusuda et al, 2002(Kusuda et al, , 2004Kobayashi et al, 2007;Yasui et al, 2011). However, the toxicity of these vehicles is species-specific in teleosts (Cabrita et al, 2006).…”