Derivation and characterization of embryonic stem‐like cells of Indian major carp Catla catla

Abstract

Embryonic stem (ES)-like cells were derived from mid-blastula stage embryos of a freshwater fish, catla Catla catla, under feeder-free condition and designated as CCES cells. The conditioned media was optimized with 10% foetal bovine serum (FBS), fish embryo extract (FEE) having 100 µg ml(-1) protein concentration, 15 ng ml(-1) basic fibroblast growth factor (bFGF) and basic media containing Leibovitz-15, DMEM with 4·5 g l(-1) glucose and Ham's F12 (LDF) in 2:1:1 ratio using a primary culture of CCES cells. Cells attached to gelatin-coated plates after 24 h of seeding and ES-like colonies were obtained at day 5 onwards. A stable cell culture was obtained after passage 10 and further maintained up to passage 44. These cells were characterized by their typical morphology, high alkaline phosphatase activity, positive expression of cell-surface antigen SSEA-1, transcription factor Oct4, germ cell marker vasa and consistent karyotype up to extended periods. The undifferentiated state was confirmed by their ability to form embryoid bodies and their differentiation potential.

Derivation and Long-Term Culture of an Embryonic Stem Cell-Like Line from Zebrafish Blastomeres Under Feeder-Free Condition

“…11,35,39 ZES1 colonies were strongly stained with AP, a common marker that has been used to positively identify pluripotency in zebrafish ES cells and other fish ES cell lines. [6][7][8][9]14,38 Our results also show how low-density culture condition induced the differentiation of ZES1 cells into neural and fibroblast cells, while muscle and epithelial cells appeared exclusively under high-density culture condition. ES cells isolated from several species, including medaka, sea bream, and sea perch, could also be directed to differentiate into neuronal or muscle-like cells when cultured in low and high density, respectively.…”

“…The medium composition was originally used to successfully derive several lineages of medaka ES cells and subsequently extended to the establishment of ES cell lines from other fish species. [6][7][8][9]11,25,35 A key difference between our tissue culture medium as compared with the previous unsuccessful formulations is the supplementation with zebrafish embryo extract. 22,26 It has been suggested that fish embryo extract contributes to the mitogenic activities of fish ES cell culture in a feeder-free culture system and could be a decisive addition in our present culture medium.…”

“…22,26 It has been suggested that fish embryo extract contributes to the mitogenic activities of fish ES cell culture in a feeder-free culture system and could be a decisive addition in our present culture medium. 6,8 The inclusion of embryo extract was also reported in the medium of the feeder-free ES cells culture protocol of several fish species. 6,7,15,36 The attributes of pluripotent mouse ES cells include stable long-term culture, small and polygonal cell morphology, normal karyotype, high AP activity, and the capacity to differentiate into specific cell types under defined conditions.…”

“…4 Attempts to derive ES cells from teleost species were first reported more than two decades ago and have ever since resulted in the establishment of ES-like cell lines from medaka, zebrafish, gilthead seabream, sea perch, red sea bream, Japanese flounder, Asian sea bass, turbot, Indian major carp, and Atlantic cod. [5][6][7][8][9][10][11][12][13][14][15] A common postulation on the potential application of ES cells from fish species with commercial values is the development of transgenic fish with beneficial traits. 16,17 This approach has been proposed to be more effective than the direct introduction of foreign transgenes into zygotes, which seldom produces the desired integration of the transgene.…”

Effects of the Developmental Stage of Extract Donor Embryos on the Culture of Marine Medaka Oryzias dancena Embryonic Stem Cell-like Cells

“…. , , (Catla catla; Dash et al, 2010) (Oryzias dancena; Lee et al, 2013) 7 , (Lateolabrax japonicas; Chen et al, 2003a) (Chrysophrys major; Chen et al, 2003b) 36 , (Scophtalmus maximus, Holen and Hamre, 2003) 3 . , , (Collodi and Barnes, 1990;Na et al, 2009), (Assouline et al, 2002;Maures et al, 2002;Schimmang, 2007;Hanington et al, 2008), .…”