StemSure® series
StemSure® series consists of a group of products that can support the culture of ES and iPS cells. Quality control testing is performed on each lot of these products using the mouse ES cell D3 strain.
Product Information
| Product Name | Description |
|---|---|
|
StemSure® D-MEM
|
Basal medium optimized for mouse ES cell culture. Because it does not contain L-Glutamine, it requires supplementation with L-glutamine before use.
|
|
StemSure® Serum Replacement
（SSR） |
A serum replacement for culturing ES and iPS cells. These stem cells can be stably cultured when this product is used as a substitute for serum.
|
|
StemSure® 2-Mercaptoethanol
Solution（2ME） |
A reducing reagent necessary for culturing ES and iPS cells.
|
|
StemSure® Monothioglycerol
Solution（MTG） |
A reducing reagent necessary for culturing ES and iPS cells. This is designed to replace 2ME.
|
|
StemSure® Gelatin Solution
|
A reagent for coating cell culture vessel surfaces. It is used gelatin from porcine skin.
|
|
StemSure® LIF
|
A factor essential for maintaining the undifferentiated ability of mouse ES cells.
|
|
StemSure® Freezing Medium
|
Serum-free cell freezing medium suitable for cryopreserving various animal cells, including mouse ES cells.
|
Quality Control Testing
| Product Name | Sterility Test |
pH | Osmolality | Endotoxin Test |
Mycoplasma Negative Test |
Colony Formation Assay or Cell Proliferation Assay |
ALP Staining |
Cell Viability Test |
|---|---|---|---|---|---|---|---|---|
|
D-MEM
|
○ | ○ | ○ | ○ | ○ | ○ | ○ | ― |
|
SSR
|
○ | ○ | ○ | ○ | ○ | ○ | ○ | ― |
|
2ME
|
○ | ― | ― | ― | ○ | ○ | ○ | ― |
|
MTG
|
○ | ― | ― | ― | ○ | ○ | ○ | ― |
|
Gelatin
|
○ | ― | ― | ○ | ○ | ○ | ○ | ― |
|
LIF
|
○ | ― | ― | ○ | ○ | ○ | ○ | ― |
|
Freezing Medium
|
○ | ― | ― | ○ | ○ | ― | ― | ○ |
○：Performed, －：Not Performed
Culture of Mouse ES Cells D3 strain
Cell Morphology and ALP Staining
Mouse ES cells strain grown in the StemSure® series of culture reagents formed colonies with a shiny appearance that is typical of mouse ES cells. It was confirmed thar ALP staining was positive
Cell Growth Curve
Mouse ES cells D3 strain was cultured 14 passages in each culture media indicated in the figure (below). Subsequently, the cell proliferation potential was compared. We found that the StemSure® series of culture reagents can support the growth of mouse ES cells D3 strain as efficiently as Competitor A’s products.
< Media composition >
StemSure® D-MEM + 15% SSR + 2mmol/l L-Glutamine + 1×MEM Non-essential
Amino Acids + 0.1mmol/l StemSure® 2-Mercaptoethanol or 0.5mmol/l
StemSure® Monothioglycerol + 1×Penicillin-Streptomycin +
1,000units/ml StemSure® LIF
（Cultured in 12-well plates coated with collagen）
Population Doubling Level
Subcultured using mouse ES cells D3 strain in easch culture media and examined the relationship between the number of culture days and cell population doubling level.. As shown in the figure (below), We found that the StemSure® series of culture reagents is equivalent to the population doubling level of Competitor A’s products..
< Media composition >
StemSure® D-MEM + 15% SSR + 2mmol/l L-Glutamine + 1×MEM Non-essential
Amino Acids + 0.1mmol/l StemSure® 2-Mercaptoethanol +
1×Penicillin-Streptomycin + 1,000units/ml StemSure® LIF
（Cultured in 12-well plates coated with collagen）
Expression of Undifferentiated Markers
Mouse ES cell D3 strain was cultured 8 passages in StemSure® series of culture media, and we confirmed that the undifferentiated markers (Nanog, Oct3/4, Sox2, and SSEA-1) were expressed in mouse ES cells D3 strain.
< Media composition >
StemSure® D-MEM + 15% SSR + 2mmol/l L-Glutamine + 1×MEM Non-essential Amino Acids + 0.1mmol/l StemSure® 2-Mercaptoethanol
+ 1×Penicillin-Streptomycin + 1,000units/ml StemSure® LIF
Teratoma Formation
Mouse ES Cell D3 Strain was subcultured using StemSure® series. The cells were subcutaneously injected into immunodeficient mice. We subcutaneously verified that these teratomas contained multiple cell types from each of the major cell lineages, such as neural tissue (ectoderm), cartilage (mesoderm), and luminal structures lined with ciliated epithelium (endoderm).
< Media composition >
StemSure® D-MEM + 15% SSR + 2mmol/l L-Glutamine + 1×MEM Non-essential Amino Acids + 0.1mmol/l StemSure® 2-Mercaptoethanol
+ 1×Penicillin-Streptomycin + 1,000units/ml StemSure® LIF
Culture of human iPS Cells 201B7 Strain
Colony Morphology and ALP Staining
We confirmed that StemSure® Serum Replacement (SSR) can support the growth of human iPS cell 201BH7 strain. We also verified that ALP staining for these cells was positive.
<Media composition>
D-MEM/Ham's F-12 + 20% SSR + 2mmol/l L-Glutamine +1×MEM Non-essential Amino
Acids + 0.1mmol/l StemSure® 2-Mercaptoethanol + 1×Penicillin-Streptomycin + 5ng/ml
bFGF
Expression of Undeifferentiated Markers
Human iPS cells 201B7 strain were culured in the media containing SSR, we confirmed that the undifferentiated markers (Sox2, Oct3/4, SSEA-3, SSEA-4 and Tra-1-81) were expressed in human iPS cells 201B7 strain.
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For research use or further manufacturing use only. Not for use in diagnostic procedures.
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