Details
| Original language | English |
|---|---|
| Pages (from-to) | 104-110 |
| Number of pages | 7 |
| Journal | CRYOBIOLOGY |
| Volume | 63 |
| Issue number | 2 |
| Early online date | 24 Jul 2011 |
| Publication status | Published - Oct 2011 |
Abstract
Stem cell research is a highly promising and rapidly progressing field inside regenerative medicine. Embryonic stem cells (ESCs), reprogrammed " induced pluripotent" cells (iPS), or lately protein induced pluripotent cells (piPS) share one inevitable factor: mouse embryonic feeder cells (MEFs), which are commonly used for ESC long term culture procedures and colony regeneration. These MEFs originate from different mouse strains, are inactivated by different methods and are differently cryopreserved. Incomprehensibly, there are to date no established quality control parameters for MEFs to insure consistency of ESC experiments and culture. Hence, in this work, we developed a bench-top quality control for embryonic feeder cells.According to our findings, MEFs should be inactivated by irradiation (30. Gy) and cryopreserved with optimal 10% DMSO at 1. K/min freezing velocity. Thawed cells should be free of mycoplasma and should have above 85 ± 13.1% viability. Values for the metabolic activity should be above 150 ± 10.5% and for the combined gene expression of selected marker genes above 225 ± 43.8% compared to non-irradiated, cryopreserved controls. Cells matching these criteria can be utilized for at least 12 days for ESC culture without detaching from the culture dish or disruption of the cell layer.
Keywords
- Cryopreservation, ESC, MEF, Quality control
ASJC Scopus subject areas
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In: CRYOBIOLOGY, Vol. 63, No. 2, 10.2011, p. 104-110.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Cryopreservation and quality control of mouse embryonic feeder cells
AU - Diekmann, Ulf
AU - Spindler, Ralf
AU - Wolkers, Willem F.
AU - Glasmacher, Birgit
AU - Müller, Thomas
N1 - Funding Information: Statement of funding: This work is supported by funding from the German Research Foundation (DFG) for the Cluster of Excellence REBIRTH (From Regenerative Biology to Reconstructive Therapy) and from the Institute for Transfusion Medicine of the Hannover Medical School.
PY - 2011/10
Y1 - 2011/10
N2 - Stem cell research is a highly promising and rapidly progressing field inside regenerative medicine. Embryonic stem cells (ESCs), reprogrammed " induced pluripotent" cells (iPS), or lately protein induced pluripotent cells (piPS) share one inevitable factor: mouse embryonic feeder cells (MEFs), which are commonly used for ESC long term culture procedures and colony regeneration. These MEFs originate from different mouse strains, are inactivated by different methods and are differently cryopreserved. Incomprehensibly, there are to date no established quality control parameters for MEFs to insure consistency of ESC experiments and culture. Hence, in this work, we developed a bench-top quality control for embryonic feeder cells.According to our findings, MEFs should be inactivated by irradiation (30. Gy) and cryopreserved with optimal 10% DMSO at 1. K/min freezing velocity. Thawed cells should be free of mycoplasma and should have above 85 ± 13.1% viability. Values for the metabolic activity should be above 150 ± 10.5% and for the combined gene expression of selected marker genes above 225 ± 43.8% compared to non-irradiated, cryopreserved controls. Cells matching these criteria can be utilized for at least 12 days for ESC culture without detaching from the culture dish or disruption of the cell layer.
AB - Stem cell research is a highly promising and rapidly progressing field inside regenerative medicine. Embryonic stem cells (ESCs), reprogrammed " induced pluripotent" cells (iPS), or lately protein induced pluripotent cells (piPS) share one inevitable factor: mouse embryonic feeder cells (MEFs), which are commonly used for ESC long term culture procedures and colony regeneration. These MEFs originate from different mouse strains, are inactivated by different methods and are differently cryopreserved. Incomprehensibly, there are to date no established quality control parameters for MEFs to insure consistency of ESC experiments and culture. Hence, in this work, we developed a bench-top quality control for embryonic feeder cells.According to our findings, MEFs should be inactivated by irradiation (30. Gy) and cryopreserved with optimal 10% DMSO at 1. K/min freezing velocity. Thawed cells should be free of mycoplasma and should have above 85 ± 13.1% viability. Values for the metabolic activity should be above 150 ± 10.5% and for the combined gene expression of selected marker genes above 225 ± 43.8% compared to non-irradiated, cryopreserved controls. Cells matching these criteria can be utilized for at least 12 days for ESC culture without detaching from the culture dish or disruption of the cell layer.
KW - Cryopreservation
KW - ESC
KW - MEF
KW - Quality control
UR - http://www.scopus.com/inward/record.url?scp=81155161943&partnerID=8YFLogxK
U2 - 10.1016/j.cryobiol.2011.07.002
DO - 10.1016/j.cryobiol.2011.07.002
M3 - Article
C2 - 21810414
AN - SCOPUS:81155161943
VL - 63
SP - 104
EP - 110
JO - CRYOBIOLOGY
JF - CRYOBIOLOGY
SN - 0011-2240
IS - 2
ER -