NDLI: Towards obtaining long term embryonic stem cell like cultures from a marine flatfish, Scophtalmus maximus

Content Provider	Springer Nature Link
Author	Holen, E. Hamre, K.
Copyright Year	2003
Abstract	Culturing pluripotent embryonic stem cells represents a unique model system for in vitro studies of embryo cell growth and differentiation, and represents a connection between in vitro and in vivo manipulation of genes. To further develop and refine stem cell technology for marine fish, we have established cultures of embryonic stem cells isolated from turbot blastulas. The pluripotent nature of our turbot-ES-like cells was supported by their morphology and elevated levels of alkaline phosphatase enzyme activity, their ability to remain undifferentiated for a prolonged culture period, their spontaneous differentiation potential in vitro and their ability to form embryoid bodies (EB) in response to changes in the extracellular environment. In addition, we show that turbot ES like cells express Oct-4 required for the maintenance of pluripotency of ES cells.Cells from 100 blastulas (>105 cells/well) were seeded into gelatine coated 24 well cell culture clusters. The cells were polygonal in shape, with dense cytoplasm and large nuclei. The ES-like cells formed colonies within 24 h following seeding, multilayered in a pyramidal fashion, with maximum cell densities in the middle. The cells proliferated vigorously when seeding densities were high and the cells still had not attached to the gelatine-coated surface. Most of the cells became attached to the surface 48 h following seeding. Attached cells grew more slowly and 20% of the plated colonies could be kept stable for 60 days. Eventually, most of the cultures showed extensive differentiation or died. Only a few cultures (4–5%) survived prolonged culturing (>2 months). The cells were stained for alkaline phosphatase activity, a marker of pluripotency and showed intense staining. More specific, turbot ES like cells in culture expressed Oct-4, detected by immunofluorescence staining.Changing the medium conditions by adding retinoic acid and removing LIF, the proportion of embryoid bodies in our cultures increased. ES-like cells as well as fresh, intact fertilised eggs where successfully cryopreserved. ES cells from the cryopreserved eggs could be isolated and seeded into cultures, forming colonies like the cells from freshly fertilised eggs. Also cryopreserved ES-like cells could be successfully plated. The prolonged survival of these cryopreserved cells has not yet been investigated. The establishment of in vitro cultures of turbot ES-like cells represents a new experimental model for marine flatfish.
Starting Page	245
Ending Page	252
Page Count	8
File Format	PDF
ISSN	09201742
Journal	Fish Physiology and Biochemistry
Volume Number	29
Issue Number	3
e-ISSN	15735168
Language	English
Publisher	Kluwer Academic Publishers
Publisher Date	2003-01-01
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Animal Biochemistry Hydrobiology Zoology Animal Anatomy / Morphology / Histology Animal Physiology
Content Type	Text
Resource Type	Article
Subject	Physiology Medicine Biochemistry Aquatic Science

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Identification of embryonic stem cell activities in an embryonic cell line derived from marine medaka (Oryzias dancena)

Embryonic stem cells in fish: current status and perspectives

Effects of hypoxia on respiratory physiology of turbot, Scophthalmus maximus

Apoptosis during gametogenesis in diploid and triploid turbot (Scophthalmus maximus)

Purification and characterization of stomach protease from the turbot (Scophthalmus maximus L.)

Development and characterization of a new marine fish cell line from turbot (Scophthalmus maximus)

Protein kinase C in the spleen of the turbot (Scophthalmus maximus L.)

Development of an ES-like cell culture system (RESC) from rohu, Labeo rohita (Ham.)

Effect of an immunostimulating alginate on protein turnover of turbot (Scophthalmus maximus L.) larvae

Towards obtaining long term embryonic stem cell like cultures from a marine flatfish, Scophtalmus maximus

Similar Documents

Identification of embryonic stem cell activities in an embryonic cell line derived from marine medaka (Oryzias dancena)

Embryonic stem cells in fish: current status and perspectives

Effects of hypoxia on respiratory physiology of turbot, Scophthalmus maximus

Apoptosis during gametogenesis in diploid and triploid turbot (Scophthalmus maximus)

Purification and characterization of stomach protease from the turbot (Scophthalmus maximus L.)

Development and characterization of a new marine fish cell line from turbot (Scophthalmus maximus)

Protein kinase C in the spleen of the turbot (Scophthalmus maximus L.)

Development of an ES-like cell culture system (RESC) from rohu, Labeo rohita (Ham.)

Effect of an immunostimulating alginate on protein turnover of turbot (Scophthalmus maximus L.) larvae

Towards obtaining long term embryonic stem cell like cultures from a marine flatfish, Scophtalmus maximus