Thermo Scientific Nunc UpCell Surface

Thermo Scientific Nunc UpCell Surface

UpCell surface is a temperature-responsive cell culture surface that enables non-enzymatic harvesting of adherent cells for preservation of cell viability and surface proteins.

Simply transfer the culture vessel to room temperature to collect cells in suspension or in the form of "cell sheet" with the help of a supportive membrane.

Nunc UpCell Surface Features:

  • No trypsin—preserves cell surface proteins
  • No scraping—promotes improved cell viability
  • Designed for culture passaging, single-cell analyses and cell transplantation research
  • The supportive membrane option enables harvesting of cell sheets and creations of 3D tissue models
  • 3 x 3mm grid design in the 60mm and 100mm dishes helps prevent cells from clumping and enables harvesting of a single-cell suspension

How does UpCell surface work?

A temperature-responsive poly(N-isopropylacrylamide) (PNIPAAm) polymer is grafted onto the standard cell culture treated polystyrene surface. At 37˚C, the covalently bound polymer allows cells to attach to the dish and grow as normal. At room temperature, the PNIPAAm binds water and swells, releasing the attached cells together with the extracellular matrices. The cells are dissociated from the culture substrate without trypsin.

Illustration showing cell adhesion at 37C and cell detachment at room temperature of the Nunc UpCell surface

Nunc UpCell surface applications

Preserve surface membrane proteins

Enzymatic cell detachment compromises the integrity of cell surface proteins. The Nunc UpCell surface is designed to offer maximum preservation of cell-surface proteins.

Flow cytometry histograms of cell number as a function of fluorescence intensity for trypsinized cells compared to those grown on Nunc UpCell surface
Figure: Shifting of the gray peaks towards right indicates that the cell surface molecule CD140a is preserved by temperature-reduction harvesting on the Nunc UpCell Surface.

Cell sheet harvesting and engineering

Illustration showing the basic steps for cultured cell sheet harvesting and engineering using Nunc UpCell surface

Cell sheets preserve cell polarization and are held together by normal cell–cell junctions and extracellular matrices. Cell sheets harvested from cultureware with Thermo Scientific Nunc UpCell Surface can be:

  • Analyzed in vitro, for example, by electron microscopy
  • Re-plated to cultureware with UpCell Surface or traditional cell cultureware
  • Used in different transplantation models
  • Stacked on top of another cell sheet for cell sheet engineering 

Cell sheet constructs prepared in cultureware with Thermo Scientific Nunc UpCell Surface can be:

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

The UpCell Surface enables harvested cell sheets to be stacked in order to form 3D tissue models. Grow your cells to confluence in cultureware with UpCell Surface, harvest the cell sheet, and transfer the cell sheet to another cell sheet. No scaffold is needed!

Examples of applications using cell sheets harvested by temperature reduction

Cell Type Application Reference
Aortic endothelial cells (cow) Analysis (structural and matrix deposition) Kushida et al., 1999
Keratinocytes (human) Analysis (electron microscopy) Yamato et al., 2001
Urothelial cells (human) Analysis (electron microscopy) Shirayanagi 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 & analysis (electron and fl uorescence microscopy) Kushida et al., 2005
Lung cells (rat) Re-plating to traditional cultureware & analysis (fl uorescence microscopy) Nandkumar et al., 2002
Smooth muscle cells & fi broblasts (human) Re-plating to PIPAAm surface & analysis (functional) & transplantation Hobo et al., 2008
Mesenchymal stem cells & skin fi broblasts (rat) Analysis (structural and functional) & transplantation Miyahara et al., 2006
Corneal stem cells (human and rabbit) Analysis (structural) & transplantation Nishida et al., 2004
Corneal endothelial cells (human) Analysis (structural and functional) & transplantation Sumide et al., 2006
Oral mucosal epithelial cells (dog) Analysis (structural) & 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

Examples of cell sheet engineering using cell sheets harvested by temperature reduction

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 fi broblasts (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 2006

Harvesting highly functional APCs

Antigen-presenting cells (APCs) are a group of immune cells that mediate the cellular immune response by processing and presenting antigens for recognition by T cells. Both dendritic cells and macrophages are APCs that require attachment to the cell culture vessel to grow. Trypsinization can damage cell membranes as well as prematurely activate APCs, potentially diminishing their antigen presenting function. Nunc UpCell Surface allows for the detaching of APCs without trypsin​, generating highly viable cells with intact membrane proteins, which ultimately​ improves cellular function.

Bar charts of mean fluorescence intensity for CD86 and CD83 markers on trypsinized cells and on those grown on UpCell surface assayed
Dendritic cells harvested from Nunc UpCell Surface exhibit higher levels of surface markers. Marker protein levels assayed by flow cytometry.
Bar charts showing the percentage of detached and viable dendritic cells obtained from trypsin-EDTA detachment compared to UpCell surface detachment
Nunc UpCell Surface allows for more effective detachment of highly viable dendritic cells.
Bar charts showing viability of macrophages detached using UpCell surface temperature reduction compared to mechanical scraping and trypsinization
Gentle temperature-reduction harvest of highly viable macrophages. Mouse peritoneal macrophage cells were harvested by either by temperature reduction (for UpCell surface) or by traditional enzymatic (trypsinization) or mechanical (scraping) methods. The cells were then counted and the recovery ratio was calculated. Mean and SD is shown.

Preserve surface membrane proteins

Enzymatic cell detachment compromises the integrity of cell surface proteins. The Nunc UpCell surface is designed to offer maximum preservation of cell-surface proteins.

Flow cytometry histograms of cell number as a function of fluorescence intensity for trypsinized cells compared to those grown on Nunc UpCell surface
Figure: Shifting of the gray peaks towards right indicates that the cell surface molecule CD140a is preserved by temperature-reduction harvesting on the Nunc UpCell Surface.

Cell sheet harvesting and engineering

Illustration showing the basic steps for cultured cell sheet harvesting and engineering using Nunc UpCell surface

Cell sheets preserve cell polarization and are held together by normal cell–cell junctions and extracellular matrices. Cell sheets harvested from cultureware with Thermo Scientific Nunc UpCell Surface can be:

  • Analyzed in vitro, for example, by electron microscopy
  • Re-plated to cultureware with UpCell Surface or traditional cell cultureware
  • Used in different transplantation models
  • Stacked on top of another cell sheet for cell sheet engineering 

Cell sheet constructs prepared in cultureware with Thermo Scientific Nunc UpCell Surface can be:

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

The UpCell Surface enables harvested cell sheets to be stacked in order to form 3D tissue models. Grow your cells to confluence in cultureware with UpCell Surface, harvest the cell sheet, and transfer the cell sheet to another cell sheet. No scaffold is needed!

Examples of applications using cell sheets harvested by temperature reduction

Cell Type Application Reference
Aortic endothelial cells (cow) Analysis (structural and matrix deposition) Kushida et al., 1999
Keratinocytes (human) Analysis (electron microscopy) Yamato et al., 2001
Urothelial cells (human) Analysis (electron microscopy) Shirayanagi 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 & analysis (electron and fl uorescence microscopy) Kushida et al., 2005
Lung cells (rat) Re-plating to traditional cultureware & analysis (fl uorescence microscopy) Nandkumar et al., 2002
Smooth muscle cells & fi broblasts (human) Re-plating to PIPAAm surface & analysis (functional) & transplantation Hobo et al., 2008
Mesenchymal stem cells & skin fi broblasts (rat) Analysis (structural and functional) & transplantation Miyahara et al., 2006
Corneal stem cells (human and rabbit) Analysis (structural) & transplantation Nishida et al., 2004
Corneal endothelial cells (human) Analysis (structural and functional) & transplantation Sumide et al., 2006
Oral mucosal epithelial cells (dog) Analysis (structural) & 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

Examples of cell sheet engineering using cell sheets harvested by temperature reduction

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 fi broblasts (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 2006

Harvesting highly functional APCs

Antigen-presenting cells (APCs) are a group of immune cells that mediate the cellular immune response by processing and presenting antigens for recognition by T cells. Both dendritic cells and macrophages are APCs that require attachment to the cell culture vessel to grow. Trypsinization can damage cell membranes as well as prematurely activate APCs, potentially diminishing their antigen presenting function. Nunc UpCell Surface allows for the detaching of APCs without trypsin​, generating highly viable cells with intact membrane proteins, which ultimately​ improves cellular function.

Bar charts of mean fluorescence intensity for CD86 and CD83 markers on trypsinized cells and on those grown on UpCell surface assayed
Dendritic cells harvested from Nunc UpCell Surface exhibit higher levels of surface markers. Marker protein levels assayed by flow cytometry.
Bar charts showing the percentage of detached and viable dendritic cells obtained from trypsin-EDTA detachment compared to UpCell surface detachment
Nunc UpCell Surface allows for more effective detachment of highly viable dendritic cells.
Bar charts showing viability of macrophages detached using UpCell surface temperature reduction compared to mechanical scraping and trypsinization
Gentle temperature-reduction harvest of highly viable macrophages. Mouse peritoneal macrophage cells were harvested by either by temperature reduction (for UpCell surface) or by traditional enzymatic (trypsinization) or mechanical (scraping) methods. The cells were then counted and the recovery ratio was calculated. Mean and SD is shown.

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