PrimeSurface® Non Adherent Surface Cultureware
The PrimeSurface range of plastic culture vessels developed by the Sumitomo Bakelite Company has low cell adhesion properties due to proprietary low-protein-adsorption technology. This makes them ideal for spheroid cell colony formation.
Applications
PrimeSurface Cultureware for Stem Cell Differentiation
Embryonic stem (ES) cells are pluripotent stem cells derived from the inner cell mass of a blastocyst, formed during the early stage of embryo development a few days post fertilisation. ES cells undergo differentiation and cell specification along the three germ lineages – endoderm, ectoderm and mesoderm – which encompass all somatic cell types.
As precursors to all somatic cells, ES cells are a key focus in tissue engineering and transplantation research fields. In addition to ES cells, stem cell research also utilises induced pluripotent stem (iPS) cells. There are an artificially derived type of pluripotent stem cell produced from somatic cells by forced expression of specific genes. Similar to ES cells, iPS cells express certain stem cell genes and proteins, chromatin methylation patterns, doubling times and embryoid body (EB) formation. The size of stem cell spheroids has been reported to greatly influence stem cell differentiation and uniform EB formation is preferable.
PrimeSurface culture plates have been successfully used to induce differentiation of ES, iPS and nerve cells.
Application: Spheroid formation of ES cells
Cells: Human ES (KhES-1)
Density: 9000 cells/well
Media: GMEM+KSR+NEAA+2ME + 20µM Y-27632
Conditions: 37C with 5% CO2.
Culture period: 6 Days
PrimeSurface: MS-9096M (Conical Well)
Application: Differentitation of ES cells into Neural Retina
Spheroid formation and subsequent differentiation of human ES Cells
Cells: Human ES (KhES-1)
Density: 9000 cells/well
Media: GMEM+KSR+NEAA+2ME + 20µM Y-27632
Conditions: 37C with 5% CO2.
PrimeSurface: MS-9096V (V- Well)
A) Spheroid formation at 18h
B) Spheroid formation at day 6
C) Self-formation of retinal tissue from the aggregate of human ES Cells
Data provided by Yoshiki Sasai (M.D., PhD.), Group Director, Stem Cell Technology, RIKEN Center for Developmental Biology.
Benefits
PrimeSurface products provide stable culture surfaces with a ultra-hydrophilic polymer coating containing a ‘high density –OH group’ which prevents elution of chemicals from the plate which can influence cell response and also promotes spontaneous spheroid formation within stem cell cultures. Following EB formation the ES cells can be stimulated to induce cell differentiation.
Uniform Spheroid Formation
Spheroids collected from seven culture wells after 6 days showing consistent size
Data provided by Yoshiki Sasai (M.D., PhD.), Group Director, Stem Cell Technology, RIKEN Center for Developmental Biology.
Evaluation and Product Comparison
To evaluate the suitability of plates for uniform spheroid formation the PrimeSurface U well plates were compared with competitor products:
| Manufacturer | Products | Grade | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Manufacturer | Products | Grade | A | B | C | D | |||||
| Manufacturer | Products | Grade | Sumitomo Co. Ltd | PrimeSurface MS-9096U | 95 | 1 | 0 | 0 | |||
| Manufacturer | Products | Grade | Company X | Product X | 87 | 6 | 3 | 0 | |||
| Manufacturer | Products | Grade | Company Y | Product Y | 83 | 9 | 4 | 0 | |||
| Manufacturer | Products | Grade | Company Z | Product Z | 0 | 0 | 49 | 47 | |||
Cells: HepG2 cells
Density: 1000 cells/100µl/Well)
Conditions: DMEM low glucose + 10% FCS
Culture Period: 3 days
The performance was graded from A to D; A was indicative of a single spheroid, B-C multiple spheroids of various sizes and D cell adhesion failure to generated cell spheroid.
PrimeSurface delivered the best performance delivering a single spheroid in 95 out of 96 wells.
Additional Information
Time Course: Mouse ES Spheroid (Embryoid Body) Formation
Cells: Mouse ES Cells 129SV
Density: 1500 cells/well
Media: DMEM + 4.5 mg/ml Glc + 15% 9v/v) heat inactivated FCS +1% (v/v) NEAA + L-Glutamine (2mM) + b-Mercaptoethanol (110 µM/ml) + 1% (V/V) Pen.-Strep
Conditions: 37C with 5% CO2.
PrimeSurface: MS-9096M Conical Well
Variety of Well Shapes for Different Applications
PrimeSurface 96 Well plates are available with different well shapes to suit different applications. Choose from U-, Conical or V- shaped wells. The well shape can influence spheroid body formation, cells with weaker aggregation properties such as the breast cancer cells MDA-MB-468 benefit from the V-shaped wells (see below) but other cultures will favour different well shapes for example mouse ES cell have been differentiated into cardiomyocytes using U-shaped wells (Yasuda et al, 2011*).
Cells: Breast cancer MDA-MD-453 & MDA-MB-468
Density: 2x103 cells/well
Media: RPMI + 10% FBS
Conditions: 37C with 5% CO2
Culture Period: 7 Days
PrimeSurface: MS-9096U (U-Well); MS-9096M (Conical Well); MS-9096V (V-Well)
Data provided by Nishio Lab., Dept. of Genome Bio. Kinki Univ. Faculty of Medicine
