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Nunc UpCell Non-Enzymatic Surfaces for Adherent Cell Culture
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
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).
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
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.
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
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:
- Seed cells in your UpCell surface-treated cultureware
- Plate at 37°C
- Grow cells up to 70% confluence (depending on cell type)
- Place cultureware in 20–25°C environment for 10–40 minutes (depending on cell type)
- And harvest your cells!
Citations for single-cell suspension harvesting with UpCell surfaces
| Cell type | Application | Reference |
|---|---|---|
| Microglia (rat) | Analysis (detachment and function) | Nakajima et al., 2001 |
| Monocytes and macrophages (human) | Re-seeding/passaging | Collier 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 resonance | Yanase 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:
- Grow cells to confluence on UpCell surface
- Aspirate/add fresh medium
- Place provided membrane on top of cell layer
- Place cultureware in 20–25°C environment for 5–6 minutes
- Manually detach cell layer around membrane edge
- Withdraw membrane (with cell layer)
- Transfer membrane (with cell layer) to new cultureware/graft site
- Remove membrane
Citations for cell sheet harvesting with UpCell surfaces
| Cell type | Application | Reference |
|---|---|---|
| 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 transplantation | Miyahara et al., 2006 |
| Corneal endothelial cells (human) | Analysis (structural and functional) and transplantation | Sumide 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:
- Transfer UpCell surface–harvested membrane (with cell sheet) to a confluent culture with aspirated medium
- Add fresh medium
- Place cultureware in a 37°C environment for ~30 minutes
- Withdraw membrane (without cell layer)
- Add fresh medium and incubate at 37°C
OR
Aspirate medium and transfer another cell sheet to construct.
Citations for cell sheet engineering with UpCell surfaces
| Cell type | Application | Reference |
|---|---|---|
| Aortic endothelial cells (human) | Cultivation (3D co-culture) & analysis (structural) | Harimoto et al., 2002 |
| Hepatocytes (mouse and human) | Analysis (structural and functional) & stacking during transplantation | Ohashi et al., 2007 |
| Skeletal myoblast (dog) | Analysis (structural) & stacking during transplantation | Hata et al., 2006 |
| Lung and skin fibroblasts (rat) | Analysis (structural) & stacking during transplantation | Kanzaki et al., 2007 |
| Cardiomyocytes (rat) | Cultivation & analysis (structural and functional) & stacking before transplantation | Sekiya 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
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:
- Seed cells in your UpCell surface-treated cultureware
- Plate at 37°C
- Grow cells up to 70% confluence (depending on cell type)
- Place cultureware in 20–25°C environment for 10–40 minutes (depending on cell type)
- And harvest your cells!
Citations for single-cell suspension harvesting with UpCell surfaces
| Cell type | Application | Reference |
|---|---|---|
| Microglia (rat) | Analysis (detachment and function) | Nakajima et al., 2001 |
| Monocytes and macrophages (human) | Re-seeding/passaging | Collier 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 resonance | Yanase 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:
- Grow cells to confluence on UpCell surface
- Aspirate/add fresh medium
- Place provided membrane on top of cell layer
- Place cultureware in 20–25°C environment for 5–6 minutes
- Manually detach cell layer around membrane edge
- Withdraw membrane (with cell layer)
- Transfer membrane (with cell layer) to new cultureware/graft site
- Remove membrane
Citations for cell sheet harvesting with UpCell surfaces
| Cell type | Application | Reference |
|---|---|---|
| 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 transplantation | Miyahara et al., 2006 |
| Corneal endothelial cells (human) | Analysis (structural and functional) and transplantation | Sumide 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:
- Transfer UpCell surface–harvested membrane (with cell sheet) to a confluent culture with aspirated medium
- Add fresh medium
- Place cultureware in a 37°C environment for ~30 minutes
- Withdraw membrane (without cell layer)
- Add fresh medium and incubate at 37°C
OR
Aspirate medium and transfer another cell sheet to construct.
Citations for cell sheet engineering with UpCell surfaces
| Cell type | Application | Reference |
|---|---|---|
| Aortic endothelial cells (human) | Cultivation (3D co-culture) & analysis (structural) | Harimoto et al., 2002 |
| Hepatocytes (mouse and human) | Analysis (structural and functional) & stacking during transplantation | Ohashi et al., 2007 |
| Skeletal myoblast (dog) | Analysis (structural) & stacking during transplantation | Hata et al., 2006 |
| Lung and skin fibroblasts (rat) | Analysis (structural) & stacking during transplantation | Kanzaki et al., 2007 |
| Cardiomyocytes (rat) | Cultivation & analysis (structural and functional) & stacking before transplantation | Sekiya 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).
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