Cryopreservation of human embryonic stem cells by a programmed freezer with an oscillating magnetic field

Pei Yi Lin, Yao Chen Yang, Shih Han Hung, Sheng Yang Lee, Maw Sheng Lee, I. Ming Chu, Shiaw Min Hwang

研究成果: 雜誌貢獻文章

17 引文 (Scopus)

摘要

Human embryonic stem cells (hESCs), due to their self-renewal capacity and pluripotency, are an important source of cells for regenerative medicine. The immediate obstacles that need to be addressed are the poor cell survival rate of hESCs and their cell quality after cryopreservation. In this study, we used the Cell Alive System (CAS) which combines a programmed freezer with an oscillating magnetic field to reduce cryo-injury during the freezing process. The hESC clumps suspended in freezing medium were divided into three groups: (i) cells frozen by a conventional freezing container, Mr. Frosty and kept in a -80. °C freezer (MF); (ii) cells frozen to -32. °C by CAS, and then transferred to a -80. °C freezer (CAS); (iii) cells frozen to -32. °C by CAS, and then transferred to a pre-cooled Mr. Frosty and kept in a -80. °C freezer (CAS-MF) for overnight. All cryovials were placed in liquid nitrogen for one week, and hESCs were then thawed and cultured on feeder for 7. days. The results of alkaline phosphatase (AP) staining showed that the attachment efficiency of the cells cryopreserved by CAS and CAS-MF was significantly higher (29.0% and 44.0%) than in the MF method (7.0%). Furthermore, we confirmed the cells cryopreserved using CAS-MF could be subcultured while expressing pluripotent markers, differentiate into three germ layers, and maintain a normal karyotype. These results demonstrate that the use of CAS-MF offers an efficient method of hESC banking.
原文英語
頁(從 - 到)256-260
頁數5
期刊Cryobiology
66
發行號3
DOIs
出版狀態已發佈 - 六月 2013

指紋

freezers
Cryopreservation
embryonic stem cells
Magnetic Fields
magnetic fields
Stem cells
cryopreservation
Magnetic fields
Freezing
cells
Liquid nitrogen
Containers
Alkaline Phosphatase
freezing
Cells
Human Embryonic Stem Cells
Germ Layers
banking
Regenerative Medicine
Karyotype

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

引用此文

Cryopreservation of human embryonic stem cells by a programmed freezer with an oscillating magnetic field. / Lin, Pei Yi; Yang, Yao Chen; Hung, Shih Han; Lee, Sheng Yang; Lee, Maw Sheng; Chu, I. Ming; Hwang, Shiaw Min.

於: Cryobiology, 卷 66, 編號 3, 06.2013, p. 256-260.

研究成果: 雜誌貢獻文章

Lin, Pei Yi ; Yang, Yao Chen ; Hung, Shih Han ; Lee, Sheng Yang ; Lee, Maw Sheng ; Chu, I. Ming ; Hwang, Shiaw Min. / Cryopreservation of human embryonic stem cells by a programmed freezer with an oscillating magnetic field. 於: Cryobiology. 2013 ; 卷 66, 編號 3. 頁 256-260.
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abstract = "Human embryonic stem cells (hESCs), due to their self-renewal capacity and pluripotency, are an important source of cells for regenerative medicine. The immediate obstacles that need to be addressed are the poor cell survival rate of hESCs and their cell quality after cryopreservation. In this study, we used the Cell Alive System (CAS) which combines a programmed freezer with an oscillating magnetic field to reduce cryo-injury during the freezing process. The hESC clumps suspended in freezing medium were divided into three groups: (i) cells frozen by a conventional freezing container, Mr. Frosty and kept in a -80. °C freezer (MF); (ii) cells frozen to -32. °C by CAS, and then transferred to a -80. °C freezer (CAS); (iii) cells frozen to -32. °C by CAS, and then transferred to a pre-cooled Mr. Frosty and kept in a -80. °C freezer (CAS-MF) for overnight. All cryovials were placed in liquid nitrogen for one week, and hESCs were then thawed and cultured on feeder for 7. days. The results of alkaline phosphatase (AP) staining showed that the attachment efficiency of the cells cryopreserved by CAS and CAS-MF was significantly higher (29.0{\%} and 44.0{\%}) than in the MF method (7.0{\%}). Furthermore, we confirmed the cells cryopreserved using CAS-MF could be subcultured while expressing pluripotent markers, differentiate into three germ layers, and maintain a normal karyotype. These results demonstrate that the use of CAS-MF offers an efficient method of hESC banking.",
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AU - Chu, I. Ming

AU - Hwang, Shiaw Min

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AB - Human embryonic stem cells (hESCs), due to their self-renewal capacity and pluripotency, are an important source of cells for regenerative medicine. The immediate obstacles that need to be addressed are the poor cell survival rate of hESCs and their cell quality after cryopreservation. In this study, we used the Cell Alive System (CAS) which combines a programmed freezer with an oscillating magnetic field to reduce cryo-injury during the freezing process. The hESC clumps suspended in freezing medium were divided into three groups: (i) cells frozen by a conventional freezing container, Mr. Frosty and kept in a -80. °C freezer (MF); (ii) cells frozen to -32. °C by CAS, and then transferred to a -80. °C freezer (CAS); (iii) cells frozen to -32. °C by CAS, and then transferred to a pre-cooled Mr. Frosty and kept in a -80. °C freezer (CAS-MF) for overnight. All cryovials were placed in liquid nitrogen for one week, and hESCs were then thawed and cultured on feeder for 7. days. The results of alkaline phosphatase (AP) staining showed that the attachment efficiency of the cells cryopreserved by CAS and CAS-MF was significantly higher (29.0% and 44.0%) than in the MF method (7.0%). Furthermore, we confirmed the cells cryopreserved using CAS-MF could be subcultured while expressing pluripotent markers, differentiate into three germ layers, and maintain a normal karyotype. These results demonstrate that the use of CAS-MF offers an efficient method of hESC banking.

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