Establishment of a Coilia nasus Gonadal Somatic Cell Line Capable of Sperm Induction In Vitro

Abstract

Coilia nasus is an important economic anadromous migratory fish of the Yangtze River in China. In recent years, overfishing and the deterioration of the ecological environment almost led to the extinction of the wild resources of C.nasus. Thus, there is an urgent need to protect this endangered fish. Recently, cell lines derived from fish have proven a promising tool for studying important aspects of aquaculture. In this study, a stable C. nasus gonadal somatic cell line (CnCSC) was established and characterized. After over one year of cell culture (>80 passages), this cell line kept stable growth. RT-PCR results revealed that the CnGSC expressed some somatic cell markers such as clu, fshr, hsd3β, and sox9b instead of germ cell markers like dazl, piwi, and vasa. The strong phagocytic activity of CnGSC suggested that it contained a large number of Sertoli cells. Interestingly, CnGSC could induce medaka spermatogonial cells (SG3) to differentiate into elongated spermatids while co-cultured together. In conclusion, we established a C. nasus gonadal somatic cell line capable of sperm induction in vitro. This research provides scientific evidence for the long-term culture of a gonadal cell line from farmed fish, which would lay the foundation for exploring the regulatory mechanisms between germ cells and somatic cells in fish.

Generation of a Normal Long-Term-Cultured Chinese Hook Snout Carp Spermatogonial Stem Cell Line Capable of Sperm Production In Vitro

“…Previous studies have shown that in lower vertebrates, spermatogenesis can be performed in vitro. In medaka, coculture of the SG3 cell line with gonadal somatic cells from rainbow trout or Coilia nasus [ 47 ] can induce sperm in vitro [ 16 ]. Recently, the whole spermatogenesis process of mice was reproduced in vitro by combining IPS induction technology and 3D culture technology, and the generated organoids formed a structure similar to seminiferous tubules, in which spermatogenesis was performed and functional sperm was obtained [ 10 , 11 ].…”

A delayed and unsynchronized ovary development as revealed by transcriptome of brain and pituitary of Coilia nasus

“…In addition, the biological signaling pathways are involved in steroidogenesis to mediate the gonad development of teleost fish, such as estrogen and TGF-β signaling pathway ( Baroiller and Guiguen, 2001 ; Amberg et al, 2013 ). Recently data from taper-tail anchovy showed that several genes involved in the sex differentiation cascade exhibited characteristic expression in both gonadal somatic cell lines cultured in vitro ( Kan et al, 2022 ) and in spermatogonial stem cell lines under long-term in vitro culture conditions ( Gu et al, 2023 ).…”

Establishment of a Coilia nasus Spermatogonial Stem Cell Line Capable of Spermatogenesis In Vitro

“…Very little research progress has been made on its artificial breeding [ 17 ]. Up to now, the gonadal somatic cell line CnGSC has been isolated from C. nasus [ 1 ]. In the current research, we successfully established the C. nasus spermatogonial cell line (CnSSC) and investigated the in vitro sperm production capability of CnSSC with gonadal somatic cells induction.…”

“…Fish species are the most abundant of all vertebrates, but several are in danger of extinction due to severe population decreases [1]. As a consequence, the conservation Biology 2023, 12, 1175 2 of 12 study of fish germplasm resources is becoming more and more crucial to the preservation of fish in danger.…”