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Chromosomal Spread Preparation of Human Embryonic Stem Cells for Karyotyping

By Priscila B. Campos, Rafaela C. Sartore, Stacie N. Abdalla and Stevens K. Rehen

Abstract

Although human embryonic stem cells (hESC) have been shown to present a stable diploid karyotype 1, many studies have reported that depending on culture conditions they become prone to acquire chromosomal anomalies such as addition of whole or parts of chromosomes. Indeed, during long-term culture, karyotypic alterations are observed when enzymatic or chemical dissociation are used 2,3,4, while manual dissection of colonies for passaging retains a stable karyotype 5. Besides, changes in the environment such as the removal of feeder cells also seem to compromise the genetic integrity of hESC 3,6. Once chromosomal alterations could affect cellular physiology, the characterization of the genetic integrity of hESC invitro is crucial considering hESC as an essential tool in embryogenesis studies and drug testing. Furthermore, for future therapeutic purposes chromosomal changes are a real concern as it is frequently associated to carcinogenesis

Topics: Cellular Biology
Publisher: MyJove Corporation
OAI identifier: oai:pubmedcentral.nih.gov:3150060
Provided by: PubMed Central
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