Cryopreservation of specialized chicken lines using cultured primordial germ cells

Abstract

Biosecurity and sustainability in poultry production requires reliable germplasm conservation. Germplasm conservation in poultry is more challenging in comparison to other livestock species. Embryo cryopreservation is not feasible for egg-laying animals, and chicken semen conservation has variable success for different chicken breeds. A potential solution is the cryopreservation of the committed diploid stem cell precursors to the gametes, the primordial germ cells (PGCs). Primordial germ cells are the lineage-restricted cells found at early embryonic stages in birds and form the sperm and eggs. We demonstrate here, using flocks of partially inbred, lower-fertility, major histocompatibility complex- (MHC-) restricted lines of chicken, that we can easily derive and cryopreserve a sufficient number of independent lines of male and female PGCs that would be sufficient to reconstitute a poultry breed. We demonstrate that germ-line transmission can be attained from these PGCs using a commercial layer line of chickens as a surrogate host. This research is a major step in developing and demonstrating that cryopreserved PGCs could be used for the biobanking of specialized flocks of birds used in research settings. The prospective application of this technology to poultry production will further increase sustainability to meet current and future production needs.

Poultry genetic resource conservation using primordial germ cells

“…Furthermore, a lower osmolality condition (250 mOsm/kg), one of the characteristics of the defined culture system, also enabled efficient derivation and propagation of both male and female chicken PGCs. Establishment of long-term culture systems of chicken PGCs provides the opportunity to significantly amplify donor PGCs before cryopreservation [51] as well as to manipulate the genome with subsequent cloning [52,53,54] in a manner similar to mouse embryonic stem (ES) cells in the chicken. Further investigations of culture conditions of non-chicken avian PGCs may help to conserve the genetic resources of wild birds ex situ .…”

Effect of crocin and naringenin supplementation in cryopreservation medium on post-thawed rooster sperm quality and expression of apoptosis associated genes

“…Avian sperm are particularly susceptible to oxidative stress [6], though reactive oxygen species (ROS), in physiological quantities, are necessary for important sperm events leading to successful fertilization [7]. In sperm, oxidative stress disturb motility and mitochondrial activity [8]; induces lipid peroxidation of the membrane [9]; and the oxidation and DNA fragmentation [10].…”

The current state of the problem of in vitrogene pool preservation in poultry

“…PGCs act as a valuable source material for cellular genetic engineering, germ plasm production, and genetic conservation of species and populations (Kino et al, 1997). Indeed, bird PGCs can be reproduced in culture and conserved without irreversible alteration of their biological properties (van der Lavoir et al, 2006;Nandi et al, 2016;Tonus et al, 2016). Two main methods are commonly used for cryopreservation of PGCs in birds: slow freezing (SLF) and ultrafast freezing (Vitrif) (Tonus et al, 2017).…”