Two lidded-12 well plates with Nunc UpCell surface
Three plates at different temperatures. At 37°C, cells adhere to the dish, at 20°C, cells are available for harvesting

Why use UpCell dishes for adherent cell culture?

With UpCell surfaces:

  • There’s no need for trypsin—UpCell non-enzymatic surfaces mean better preservation of cell surface proteins
  • There’s no need for scraping—UpCell surfaces promote improved cell viability
  • Plastics are designed for culture passaging, single-cell analyses, and cell transplantation research
  • With supportive membrane options, UpCell surfaces enable harvesting of cell sheets and the creation of 3D tissue models

UpCell surfaces and trypsin: cell viability

Despite longer dissociation time, UpCell surfaces exhibit minimal impact on cell viability. These right-hand graphs show mesenchymal stem cells (MSCs) differential viability retention when MSCs, treated with either trypsin on the Nunclon Delta Surface or dissociated by temperature shift on the Nunc UpCell surface, were left until completely detached.

(A) Dissociation time was noted.
(B) Cell viability was measured using the Countess II Automated Cell Counter.

Please note: Individual experiments were done in duplicate, and data are represented as the mean ± SEM. ns: not significant, *: P < 0.05 (two-tailed unpaired t-test)

 Explore more comparative results between trypsinization and UpCell

A bar chart in reds describing Nunc UpCell surface’s longer cell dissociation time
A bar chart in reds describing Nunc UpCell surface’s superior ability for cell viability retention even as harvesting takes longer by UpCell surface.

UpCell surfaces and trypsin: protein expression

With measurements of CD44, CD105, and CD13 fluorescence intensity in MSCs after antibody staining, results indicate that UpCell surfaces have milder effects on cultured MSCs, preserving cell-surface antigens and maintaining high viability when compared to trypsinization.

(C) Representative flow cytometry histograms showing expression of CD44 following dissociation using 0.25% trypsin or the Nunc UpCell Surface.
(D) Median fluorescence intensities of surface antigens are demonstrated as bar graphs.

Please note: Individual experiments were done in duplicate, and data are represented as the mean ± SEM. ns: not significant, **: P < 0.005, *: P < 0.05 (two-tailed unpaired t-test).

 Explore comparative results for CD105 and CD13

Histogram of count vs. log fluorescence intensity for CD44 staining following trypsinization vs UpCell surface
A log fluorescence intensity assay describing superior retention of CD44 antigens in MSCs after dissociation by UpCell Surface.

UpCell surfaces and scraping: cell recovery

Recovery ratios of mouse peritoneal macrophages harvested from the Nunc UpCell surfaces were compared with recovery ratios of these cells harvested by either enzymatic (trypsinization) or mechanical (scraping) methods.

The recovery of cells from the Nunc UpCell surface was significantly higher than the recovery of cells harvested from traditional cultureware by trypsinization or scraping (see figure).

Please note: Mean and SD are shown.

 Explore more comparative results between cell scraping and UpCell

A bar chart of recovery ratio for UpCell surface compared to trypsinization or cell scraping

How the Nunc UpCell surface works

The Nunc UpCell surface is designed to be temperature responsive, enabling non-enzymatic cell dissociation. This surface is composed of a covalently immobilized polymer, PIPAAm. PIPAAm forms a thin, even coating over the cultureware and, being slightly hydrophobic at 37°C, allows cells to attach and grow. However, when the cultureware’s temperature reduces below 32°C, this same layer becomes hydrophilic. In this state, PIPAAm binds water, swells, and results in adherent cell release.

Order Nunc UpCell surface cultureware

At 37°C, UpCell surfaces are hydrophobic, allowing cell adhesion, below 32°C, the UpCell surface becomes hydrophilic, binds water, swells, and releases cells.


Nunc UpCell surface applications

Maximize cell-surface protein preservation

While many surface proteins are critical for the cell’s response to extracellular matrices, to other cells, and to growth factors, as well as other soluble mediators, enzymatic cell harvesting such as trypsinization compromises these proteins’ integrity.

The right-shifting of gray peaks (see figure) indicate that CD140a, a cell-surface molecule, is preserved through temperature-reduction harvesting on the Nunc UpCell surface as compared with trypsinization.

 Read more about UpCell surface's cell-surface-protein preservation

: four histograms describing how CD140a, a cell surface molecule, is preserved through UpCell-surface-mediated harvesting as compared with trypsinization.

How to use UpCell surfaces for single-cell harvesting

Nunc UpCell enables high-viability harvesting, keeping cell surface receptors and antigens intact. This includes finicky cells, like primary cells, which prove difficult to passage by other methods.

Single-cell suspensions harvested from cultureware with UpCell surfaces can be utilized by: 

  • Analysis in vitro (i.e., by flow cytometry)
  • Manipulation to purify certain cell types
  • Re-seeding in cultureware with UpCell surfaces or traditional cultureware as a part of passaging procedures
  • Researching cell transplantation

To harvest single cells from UpCell surface-treated cultureware:

  1. Seed cells in your UpCell surface-treated cultureware
  2. Plate at 37°C
  3. Grow cells up to 70% confluence (depending on cell type)
  4. Place cultureware in 20–25°C environment for 10–40 minutes (depending on cell type)
  5. And harvest your cells!
Step by step instructional on how to harvest single cells with UpCell cultureware

Citations for single-cell suspension harvesting with UpCell surfaces

Cell typeApplicationReference
Microglia (rat)

Analysis (detachment and function)

Nakajima et al., 2001
Monocytes and macrophages (human)Re-seeding/passagingCollier et al., 2002
Monocytes and macrophages (human)Activation and analysis (structural)Gordon and Freedman, 2006
Basophilic cell line RBL-2H3 (rat)Antigen-mediated degranulation measured by surface plasmon resonanceYanase et al., 2007

How to use UpCell surfaces for cell-sheet harvesting

Anchorage-dependent cells are ordinarily held together by a deposited subcellular matrix as well as by cell junctions attached to medium-derived and cultureware-absorbed proteins. When using traditional enzymatic and/or mechanical harvesting methods, these cell-to-cell, and cell-to-matrix contacts, as well as the subcellular matrix itself, are disrupted—their polarization destroyed.

However, with Nunc UpCell surface and supplied support membranes, cells can be detached as contiguous sheets, avoiding this disruption. Membranes are supplied with Nunc Multidishes with UpCell Surface (Cat. No. 174901) and Nunc Dishes with UpCell Surface (Cat. No. 174904) only. 

Cell sheets harvested from UpCell surfaces can be utilized by:

  • Analysis in vitro (i.e., electron microscopy)
  • Re-plating into cultureware with UpCell surfaces or into traditional cultureware
  • Transplantation models
  • Cell sheet stacking/engineering

To harvest cell sheets from UpCell surfaces:

  1. Grow cells to confluence on UpCell surface
  2. Aspirate/add fresh medium
  3. Place provided membrane on top of cell layer
  4. Place cultureware in 20–25°C environment for 5–6 minutes
  5. Manually detach cell layer around membrane edge
  6. Withdraw membrane (with cell layer)
  7. Transfer membrane (with cell layer) to new cultureware/graft site
  8. Remove membrane
Step by step instructional on how to harvest cell sheets with UpCell cultureware

Citations for cell sheet harvesting with UpCell surfaces

Cell typeApplicationReference
Urothelial cells (human)

Analysis (electron microscopy)

Shiroyanagi et al., 2003 
Retinal pigment epithelial cell line ARPE-19 (human)Analysis (light microscopy)Kubota et al., 2006 
Kidney epithelial cells (human and dog)Re-plating to traditional cultureware and analysis (electron and fluorescence microscopy)Kushida et al., 2005 
Mesenchymal stem cells and skin fibroblasts (rat)Analysis (structural and functional) and transplantationMiyahara et al., 2006 
Corneal endothelial cells (human)Analysis (structural and functional) and transplantationSumide et al., 2006 
Oral mucosal epithelial cells (dog)

Analysis (structural) and transplantation

Ohki et al., 2006 
Tracheal epithelial cells (rabbit)

Transplantation

Kanzaki et al., 2006 
Periodontal ligament cells (human)

Transplantation

Hasegawa et al., 2005 
Mesenchymal stem cells (human)

Transplantation

Cho et al., 2020 

How to use UpCell surfaces for 3D tissue models

Ordinarily, 3D tissue models or transplants are prepared by seeding cell suspensions on prefabricated scaffolds. These scaffold’s materials are not ordinarily cell produced. Instead, foreign, or xenogeneic materials, such as polylactic acid (PLA), polyglycolic acid (PGA), alginate, gelatin, and collagen are often utilized.

With this procedure, problems such as uneven cell distribution, special distribution control difficulties, host inflammatory reactions, or fibrous tissue formation can arise.

To mitigate these risks, UpCell surfaces enable harvested cell sheets to be stacked to form 3D tissue models—sans xenogeneic scaffold.

These Nunc UpCell surface cell sheet constructs can be:

  • Analyzed in vitro (i.e., by functional tissue-specific tests)
  • Cultivated in vitro (i.e., as 3D co-cultures)
  • Used in different transplantation models, where cell sheets are stacked before or during the transplantation procedure

To engineer cell sheets with UpCell surfaces:

  1. Transfer UpCell surface–harvested membrane (with cell sheet) to a confluent culture with aspirated medium
  2. Add fresh medium
  3. Place cultureware in a 37°C environment for ~30 minutes
  4. Withdraw membrane (without cell layer)
  5. Add fresh medium and incubate at 37°C

OR

Aspirate medium and transfer another cell sheet to construct.

Step by step instructional on how to engineer cell sheets with UpCell cultureware

Citations for cell sheet engineering with UpCell surfaces

Cell typeApplicationReference
Aortic endothelial cells (human)

Cultivation (3D co-culture) & analysis (structural)

Harimoto et al., 2002 
Hepatocytes (mouse and human)Analysis (structural and functional) & stacking during transplantationOhashi et al., 2007 
Skeletal myoblast (dog)Analysis (structural) & stacking during transplantationHata et al., 2006 
Lung and skin fibroblasts (rat)Analysis (structural) & stacking during transplantationKanzaki et al., 2007 
Cardiomyocytes (rat)Cultivation & analysis (structural and functional) & stacking before transplantationSekiya et al., 2006 Shimizu et al., 2002 , and Shimizu et al., 2006 

Quality is inherent in our culture

From product development and sourcing raw materials to manufacturing and customer service, quality is reflected in every Nunc product.

A certificate of quality is packed in each box of cultureware with Nunc UpCell surface. This certificate is your confirmation that the product has been validated according to the following tests:

Cell growth

Each manufacturing lot is sampled and subjected to performance testing for growth with the 3T3-Swiss Albino cell line (derived from a mouse embryo fibroblast) in accordance with standard operating procedures.

Acceptance level: minimum of 80% confluence.

Cell detachment

The manufacturing lot is sampled and subjected to performance testing for cell detachment by temperature reduction with the 3T3-Swiss Albino cell line in accordance with standard operating procedures. Adherent cells are detached by temperature decreasing treatment (below 32°C) and the degree of detachment is measured.

Acceptance level: detachment of 50% or more of the cells.

Sterility

Sterility is obtained by using ethylene oxide gas according to ISO 11135-1 guidelines:
(Sterilization of health care products, Ethylene oxide, Part 1: Requirements for development, validation, and routine control of a sterilization process for medical devices).

Maximize cell-surface protein preservation

While many surface proteins are critical for the cell’s response to extracellular matrices, to other cells, and to growth factors, as well as other soluble mediators, enzymatic cell harvesting such as trypsinization compromises these proteins’ integrity.

The right-shifting of gray peaks (see figure) indicate that CD140a, a cell-surface molecule, is preserved through temperature-reduction harvesting on the Nunc UpCell surface as compared with trypsinization.

 Read more about UpCell surface's cell-surface-protein preservation

: four histograms describing how CD140a, a cell surface molecule, is preserved through UpCell-surface-mediated harvesting as compared with trypsinization.

How to use UpCell surfaces for single-cell harvesting

Nunc UpCell enables high-viability harvesting, keeping cell surface receptors and antigens intact. This includes finicky cells, like primary cells, which prove difficult to passage by other methods.

Single-cell suspensions harvested from cultureware with UpCell surfaces can be utilized by: 

  • Analysis in vitro (i.e., by flow cytometry)
  • Manipulation to purify certain cell types
  • Re-seeding in cultureware with UpCell surfaces or traditional cultureware as a part of passaging procedures
  • Researching cell transplantation

To harvest single cells from UpCell surface-treated cultureware:

  1. Seed cells in your UpCell surface-treated cultureware
  2. Plate at 37°C
  3. Grow cells up to 70% confluence (depending on cell type)
  4. Place cultureware in 20–25°C environment for 10–40 minutes (depending on cell type)
  5. And harvest your cells!
Step by step instructional on how to harvest single cells with UpCell cultureware

Citations for single-cell suspension harvesting with UpCell surfaces

Cell typeApplicationReference
Microglia (rat)

Analysis (detachment and function)

Nakajima et al., 2001
Monocytes and macrophages (human)Re-seeding/passagingCollier et al., 2002
Monocytes and macrophages (human)Activation and analysis (structural)Gordon and Freedman, 2006
Basophilic cell line RBL-2H3 (rat)Antigen-mediated degranulation measured by surface plasmon resonanceYanase et al., 2007

How to use UpCell surfaces for cell-sheet harvesting

Anchorage-dependent cells are ordinarily held together by a deposited subcellular matrix as well as by cell junctions attached to medium-derived and cultureware-absorbed proteins. When using traditional enzymatic and/or mechanical harvesting methods, these cell-to-cell, and cell-to-matrix contacts, as well as the subcellular matrix itself, are disrupted—their polarization destroyed.

However, with Nunc UpCell surface and supplied support membranes, cells can be detached as contiguous sheets, avoiding this disruption. Membranes are supplied with Nunc Multidishes with UpCell Surface (Cat. No. 174901) and Nunc Dishes with UpCell Surface (Cat. No. 174904) only. 

Cell sheets harvested from UpCell surfaces can be utilized by:

  • Analysis in vitro (i.e., electron microscopy)
  • Re-plating into cultureware with UpCell surfaces or into traditional cultureware
  • Transplantation models
  • Cell sheet stacking/engineering

To harvest cell sheets from UpCell surfaces:

  1. Grow cells to confluence on UpCell surface
  2. Aspirate/add fresh medium
  3. Place provided membrane on top of cell layer
  4. Place cultureware in 20–25°C environment for 5–6 minutes
  5. Manually detach cell layer around membrane edge
  6. Withdraw membrane (with cell layer)
  7. Transfer membrane (with cell layer) to new cultureware/graft site
  8. Remove membrane
Step by step instructional on how to harvest cell sheets with UpCell cultureware

Citations for cell sheet harvesting with UpCell surfaces

Cell typeApplicationReference
Urothelial cells (human)

Analysis (electron microscopy)

Shiroyanagi et al., 2003 
Retinal pigment epithelial cell line ARPE-19 (human)Analysis (light microscopy)Kubota et al., 2006 
Kidney epithelial cells (human and dog)Re-plating to traditional cultureware and analysis (electron and fluorescence microscopy)Kushida et al., 2005 
Mesenchymal stem cells and skin fibroblasts (rat)Analysis (structural and functional) and transplantationMiyahara et al., 2006 
Corneal endothelial cells (human)Analysis (structural and functional) and transplantationSumide et al., 2006 
Oral mucosal epithelial cells (dog)

Analysis (structural) and transplantation

Ohki et al., 2006 
Tracheal epithelial cells (rabbit)

Transplantation

Kanzaki et al., 2006 
Periodontal ligament cells (human)

Transplantation

Hasegawa et al., 2005 
Mesenchymal stem cells (human)

Transplantation

Cho et al., 2020 

How to use UpCell surfaces for 3D tissue models

Ordinarily, 3D tissue models or transplants are prepared by seeding cell suspensions on prefabricated scaffolds. These scaffold’s materials are not ordinarily cell produced. Instead, foreign, or xenogeneic materials, such as polylactic acid (PLA), polyglycolic acid (PGA), alginate, gelatin, and collagen are often utilized.

With this procedure, problems such as uneven cell distribution, special distribution control difficulties, host inflammatory reactions, or fibrous tissue formation can arise.

To mitigate these risks, UpCell surfaces enable harvested cell sheets to be stacked to form 3D tissue models—sans xenogeneic scaffold.

These Nunc UpCell surface cell sheet constructs can be:

  • Analyzed in vitro (i.e., by functional tissue-specific tests)
  • Cultivated in vitro (i.e., as 3D co-cultures)
  • Used in different transplantation models, where cell sheets are stacked before or during the transplantation procedure

To engineer cell sheets with UpCell surfaces:

  1. Transfer UpCell surface–harvested membrane (with cell sheet) to a confluent culture with aspirated medium
  2. Add fresh medium
  3. Place cultureware in a 37°C environment for ~30 minutes
  4. Withdraw membrane (without cell layer)
  5. Add fresh medium and incubate at 37°C

OR

Aspirate medium and transfer another cell sheet to construct.

Step by step instructional on how to engineer cell sheets with UpCell cultureware

Citations for cell sheet engineering with UpCell surfaces

Cell typeApplicationReference
Aortic endothelial cells (human)

Cultivation (3D co-culture) & analysis (structural)

Harimoto et al., 2002 
Hepatocytes (mouse and human)Analysis (structural and functional) & stacking during transplantationOhashi et al., 2007 
Skeletal myoblast (dog)Analysis (structural) & stacking during transplantationHata et al., 2006 
Lung and skin fibroblasts (rat)Analysis (structural) & stacking during transplantationKanzaki et al., 2007 
Cardiomyocytes (rat)Cultivation & analysis (structural and functional) & stacking before transplantationSekiya et al., 2006 Shimizu et al., 2002 , and Shimizu et al., 2006 

Quality is inherent in our culture

From product development and sourcing raw materials to manufacturing and customer service, quality is reflected in every Nunc product.

A certificate of quality is packed in each box of cultureware with Nunc UpCell surface. This certificate is your confirmation that the product has been validated according to the following tests:

Cell growth

Each manufacturing lot is sampled and subjected to performance testing for growth with the 3T3-Swiss Albino cell line (derived from a mouse embryo fibroblast) in accordance with standard operating procedures.

Acceptance level: minimum of 80% confluence.

Cell detachment

The manufacturing lot is sampled and subjected to performance testing for cell detachment by temperature reduction with the 3T3-Swiss Albino cell line in accordance with standard operating procedures. Adherent cells are detached by temperature decreasing treatment (below 32°C) and the degree of detachment is measured.

Acceptance level: detachment of 50% or more of the cells.

Sterility

Sterility is obtained by using ethylene oxide gas according to ISO 11135-1 guidelines:
(Sterilization of health care products, Ethylene oxide, Part 1: Requirements for development, validation, and routine control of a sterilization process for medical devices).


Order Nunc UpCell surface plates and dishes

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