JP | JPY
-
-
-
- Challenges and Chances: A Review of the 1st Stem Cell Community Day
- Summertime, and the Livin’ Is Easy…
- Follow-on-Biologics – More than Simple Generics
- Bacteria Versus Body Cells: A 1:1 Tie
- Behind the Crime Scene: How Biological Traces Can Help to Convict Offenders
- Every 3 Seconds Someone in the World Is Affected by Alzheimer's
- HIV – It’s Still Not Under Control…
- How Many Will Be Convicted This Time?
- Malaria – the Battle is Not Lost
- Physicians on Standby: The Annual Flu Season Can Be Serious
- At the Forefront in Fighting Cancer
- Molecular Motors: Think Small and yet Smaller Again…
- Liquid Biopsy: Novel Methods May Ease Cancer Detection and Therapy
- They Are Invisible, Sneaky and Disgusting – But Today It’s Their Special Day!
- How Many Cells Are in Your Body? Probably More Than You Think!
- What You Need to Know about Antibiotic Resistance – Findings, Facts and Good Intentions
- Why Do Old Men Have Big Ears?
- The Condemned Live Longer: A Potential Paradigm Shift in Genetics
- From Research to Commerce
- Chronobiology – How the Cold Seasons Influence Our Biorhythms
- Taskforce Microbots: Targeted Treatment from Inside the Body
- Eyes on Cancer Therapy
-
-
-
-
- Challenges and Chances: A Review of the 1st Stem Cell Community Day
- Summertime, and the Livin’ Is Easy…
- Follow-on-Biologics – More than Simple Generics
- Bacteria Versus Body Cells: A 1:1 Tie
- Behind the Crime Scene: How Biological Traces Can Help to Convict Offenders
- Every 3 Seconds Someone in the World Is Affected by Alzheimer's
- HIV – It’s Still Not Under Control…
- How Many Will Be Convicted This Time?
- Malaria – the Battle is Not Lost
- Physicians on Standby: The Annual Flu Season Can Be Serious
- At the Forefront in Fighting Cancer
- Molecular Motors: Think Small and yet Smaller Again…
- Liquid Biopsy: Novel Methods May Ease Cancer Detection and Therapy
- They Are Invisible, Sneaky and Disgusting – But Today It’s Their Special Day!
- How Many Cells Are in Your Body? Probably More Than You Think!
- What You Need to Know about Antibiotic Resistance – Findings, Facts and Good Intentions
- Why Do Old Men Have Big Ears?
- The Condemned Live Longer: A Potential Paradigm Shift in Genetics
- From Research to Commerce
- Chronobiology – How the Cold Seasons Influence Our Biorhythms
- Taskforce Microbots: Targeted Treatment from Inside the Body
- Eyes on Cancer Therapy
-
JP | JPY
What is the best way to thaw cells? Hand-warming vs. water bath immersion vs. automated cell thawing
Lab Academy
Ask the expert - Answer by Ines Hartmann, Application Specialist Cell Handling at Eppendorf
This article appeared first in “ Inside Cell Culture ”, the monthly newsletter for cell culture professionals.
When removing a cryovial from the precious liquid nitrogen stock to start a fresh culture, one strives for a good and reproducible cell recovery. A generally accepted method in cryopreservation is a slow freeze and rapid thaw approach [1]. Rapid thawing can be achieved by simple warming the cryovial in the hand, immersing it in a water bath or by using an appropriate thawing device. All methods result in thawing times within an acceptable range of a few minutes [2]. However, the methods have different pros and cons.
Hand-warming efficiency is strongly person-dependent. Letting three different persons thawing a cryovial by hand-warming may result in large variations in thawing time and temperature profile in the cryovial [2]. This can be explained by the difference in hand size and temperature. The method can hardly be standardized, hence, is not optimal for reproducible cell thawing.
Thawing by immersing the cryovial in a water bath is the most common and by far the fastest method among the three compared [2]. However, the water bath is a well-known source of contamination. One should make sure that the cap of the vial is tightly closed, avoid submerging the cap area and wipe the vial thoroughly with alcohol 70% prior opening it. Also, stick with one immersion practice. A change such as one time moving, one time not moving the vial in the water or other time using a floater may affect thawing times and temperature profiles, hence reproducibility [2].
A dedicated thawing device works water-free, which minimizes the risk of a contamination. Thawing occurs within an acceptable time frame and is highly reproducible [2]. Another advantage is the small footprint facilitating an easy integration into the workspace next to the biosafety cabinet. It has been shown to be suitable for cell lines as well as for sensitive stem cells [2,3].
To sum up, we recommend either using a water bath with the precautions described above or an automated cell thawing device.
References:
[1] Thompson ML, Kunkel EJ, Ehrhardt RO (2014). Cryopreservation and Thawing of Mammalian Cells. In eLS, John Wiley & Sons, Ltd (Ed.)
[2] Tacheny A, Tejerina Vargas S, Chandelier N, Hoet JF, Karow K, Hartmann I. Standardized and Water-free Cell Thawing using the Eppendorf ThermoMixer® C with the Eppendorf SmartBlock cryo thaw, APPLICATION NOTE I No. 437
[3] Tacheny A, Tejerina Vargas S, Chandelier N, Hoet JF, Vanbellinghen B, Hartmann I. Standardized and Water-free Cell Thawing of Stem Cells using the Eppendorf ThermoMixer® C with the Eppendorf SmartBlock cryo thaw, APPLICATION NOTE I No. 452
This article appeared first in “ Inside Cell Culture ”, the monthly newsletter for cell culture professionals.
When removing a cryovial from the precious liquid nitrogen stock to start a fresh culture, one strives for a good and reproducible cell recovery. A generally accepted method in cryopreservation is a slow freeze and rapid thaw approach [1]. Rapid thawing can be achieved by simple warming the cryovial in the hand, immersing it in a water bath or by using an appropriate thawing device. All methods result in thawing times within an acceptable range of a few minutes [2]. However, the methods have different pros and cons.
Hand-warming efficiency is strongly person-dependent. Letting three different persons thawing a cryovial by hand-warming may result in large variations in thawing time and temperature profile in the cryovial [2]. This can be explained by the difference in hand size and temperature. The method can hardly be standardized, hence, is not optimal for reproducible cell thawing.
Thawing by immersing the cryovial in a water bath is the most common and by far the fastest method among the three compared [2]. However, the water bath is a well-known source of contamination. One should make sure that the cap of the vial is tightly closed, avoid submerging the cap area and wipe the vial thoroughly with alcohol 70% prior opening it. Also, stick with one immersion practice. A change such as one time moving, one time not moving the vial in the water or other time using a floater may affect thawing times and temperature profiles, hence reproducibility [2].
A dedicated thawing device works water-free, which minimizes the risk of a contamination. Thawing occurs within an acceptable time frame and is highly reproducible [2]. Another advantage is the small footprint facilitating an easy integration into the workspace next to the biosafety cabinet. It has been shown to be suitable for cell lines as well as for sensitive stem cells [2,3].
To sum up, we recommend either using a water bath with the precautions described above or an automated cell thawing device.
References:
[1] Thompson ML, Kunkel EJ, Ehrhardt RO (2014). Cryopreservation and Thawing of Mammalian Cells. In eLS, John Wiley & Sons, Ltd (Ed.)
[2] Tacheny A, Tejerina Vargas S, Chandelier N, Hoet JF, Karow K, Hartmann I. Standardized and Water-free Cell Thawing using the Eppendorf ThermoMixer® C with the Eppendorf SmartBlock cryo thaw, APPLICATION NOTE I No. 437
[3] Tacheny A, Tejerina Vargas S, Chandelier N, Hoet JF, Vanbellinghen B, Hartmann I. Standardized and Water-free Cell Thawing of Stem Cells using the Eppendorf ThermoMixer® C with the Eppendorf SmartBlock cryo thaw, APPLICATION NOTE I No. 452
Read more
Read less
Related links:
Read more
Read less
Related files:
- White Paper: Cell Thawing Protocol Standardization – Guide for More Reproducible Cryo-preservation Results
Read more
Read less