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Immortalized feeders for the scale-up of human embryonic stem cells in feeder and feeder-free conditions

Paper ID Volume ID Publish Year Pages File Format Full-Text
25583 43582 2006 12 PDF Available
Title
Immortalized feeders for the scale-up of human embryonic stem cells in feeder and feeder-free conditions
Abstract

Human embryonic stem cells (hESC) are pluripotent cells that proliferate indefinitely in culture, whilst retaining their capacity for differentiation into different cell types. However, hESC cultures require culture in direct contact with feeder cells or conditioned medium (CM) from feeder cells. The most common source of feeders has been primary mouse embryonic fibroblast (MEF). In this study, we immortalized a primary MEF line with the E6 and E7 genes from HPV16. The immortal line, ΔE-MEF, was able to proliferate beyond 7–9 passages and has an extended lifespan beyond 70 passages. When tested for its ability to support hESC growth, it was found that hESC continue to maintain the undifferentiated morphology for >40 passages both in co-culture with ΔE-MEF and in feeder-free cultures supplemented with CM from ΔE-MEF. The cultures also continue to express the pluripotent markers, Oct-4, SSEA-4, Tra-1-60, Tra-1-81, alkaline phosphatase and maintain a normal karyotype. In addition, these hESC formed teratomas when injected into SCID mice. Lastly, we demonstrated the feasibility of scaling-up significant quantities of undifferentiated hESC (>108 cells) using ΔE-MEF in cell factories. The results from this study suggest that immortalized feeders can provide a consistent and reproducible source of feeders for hESC expansion and research.

Keywords
Human embryonic stem cells; Immortalized feeders; Undifferentiated
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Immortalized feeders for the scale-up of human embryonic stem cells in feeder and feeder-free conditions
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 122, Issue 1, 9 March 2006, Pages 130–141
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us