Repro-PACT™ & other new products of Cellagen Tech.
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Repro-PACT™ & other new products of Cellagen Technology LLC
Product Name: Repro-PACT™
According to literatures1-4, the supplement of small molecule cocktail (Repro-PACT) in cell culture media will simplify
the process and increase the productivity in generating iPS cells. This process can be easily scale-up simply by culturing
more cells.
Repro-PACT can be used on many cell types, including primary skin fibroblasts, peripheral blood mononuclear cells,
endothelial cells, or keratinocytes, and can be used in combination with any of the widely used reprogramming gene
expression systems (e.g. retro/lenti-viral, episomal, mRNA, or proteins) to enhance the reprogramming to iPS cells.
Details:
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Chemical Formulation: |
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DMSO solution contains: |
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1000 x DMSO stock |
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0.5 mM of PD0325901, |
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solution |
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0.5 mM of A83-01, |
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3.0 mM of CHIR99021, and |
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0.5 mM of Thiazovivin |
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Purity: |
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>98% (for each compound) |
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Shipping condition: |
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The product is shipped in a glass vial at ambient temperature. |
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Storage: |
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Store DMSO solution at -20oC. |
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Usage: |
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Dilute Repro-PACT at 1:1000 into your basic ES media to make Repro-PACT media. |
References:
1. Lin et al. (2009) A chemical platform for improve induction of human iPS cells. Nature Method 6, 805-808.
2. Kawamata et al. (2010). Generation of genetically modified rats from embryonic stem cells. PNAS 107, 14223-14228.
3. Zhu et al. (2010) Reprogramming of human primary somatic cells by OCT4 and chemical compounds. Cell Stem Cell 7,
651-655.
4. Yu et al. (2011) Efficient feeder-free episomal reprogramming with small molecules. PLoS ONE 6(3): e17557.
Product Name: Repro-ThiPSC
Repro-ThiPSC (i.e. SMC4 in reference 1) is a cocktail of four small molecules of specific signaling pathway inhibitors. It can
be added directly in media to increase reprogramming efficiency of somatic cells to iPSCs, and to improve expansion and
maintenance of hiPSCs under both with feeder cells and feeder-free conditions. It can be used in combination with any
of the widely used reprogramming gene expression systems (e.g. retro/lenti-viral, episomal, mRNA, or proteins) to
enhance the reprogramming somatic cells to iPS cells. This process is robust, efficient and can be easily scaled-up1-3.
Details:
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Chemical Formulation: |
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5 mM of Thiazovivin (ROCK inhibitor), |
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1000 x DMSO stock |
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0.4 mM of PD325901 (MEK inhibitor), |
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solution |
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2 mM of SB431542 (TGF-ß1 inhibitor), and |
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1 mM of CHIR99021 (GSK3ß inhibitor) |
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Purity: |
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>98% (for each compound) |
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Shipping condition: |
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The product is shipped in a glass vial at ambient temperature. |
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Storage: |
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Store DMSO solution at -20oC. |
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Usage: |
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Dilute Repro-ThiPSC stock solution (1000x) at 1:1000 into basic iPSC induction media |
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to make 1xRepro-ThiPSC media. |
References:
1. Valamehr B, et al. A novel platform to enable the high-throughput derivation and characterization of feeder-free
human iPSCs. Scientific Reports. 2012. 2: 213.
2. Li W, et al. Generation of human-induced pluripotent stem cells in the absence of exogenous Sox2. Stem Cells. 2009.
27(12):2992-3000.
3. Kawamata M, et al. Generation of genetically modified rats from embryonic stem cells. PNAS. 2010. 107,
14223-14228.
Product Name: Repro-nSC (1000 x)
Repro-nSC is a combination of two small molecules: a GSK-3ß inhibitor CHIR99021, and a TGF-ß receptor inhibitor SB431542. A study has shown the efficient conversion of hESCs into homogenous primitive neuroepithelium within 7 days when under
synergistic inhibition of GSK-3ß and TGF-ß pathways by Repro-nSC alone or in combination with inhibition of the
Notch signaling pathway (e.g. by RO4929097). These primitive neural stem cells can stably self-renew in the presence of LIF and 1x Repro-nSC, retaining high neurogenic potential and responsiveness to instructive neural patterning cues
towards midbrain and hindbrain neuronal subtypes, in addition to exhibiting in vivo integration1.
Details:
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Chemical Formulation: |
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2 mM of SB431542 (TGF-ß1 inhibitor) |
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1000 x DMSO stock |
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3 mM of CHIR99021 (GSK3ß inhibitor) |
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solution |
: |
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Purity: |
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>98% (for each compound) |
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Shipping condition: |
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The product is shipped in a glass vial at ambient temperature. |
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Storage: |
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Store DMSO solution at -20oC. |
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Usage: |
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Dilute Repro-nSC stock solution (1000x) at 1:1000 into your neural induction media |
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to make 1xRepro-nSC media. |
References:
1. Li W., et al. Rapid induction and long-term self-renewal of primitive neural precursors from human embryonic stem cells
by small molecule inhibitors Proc Natl Acad Sci 2011 108(20):8299-304.
Product Name: Repro-nLS3i (1000 x)
Repro-nLS3i (also named LSB3i in reference 1) contains two cocktails of total five small molecule inhibitors. According to
literature1-3, the combination of these inhibitors significantly accelerated the process of deriving postmitotic neurons
from human ESCs and iPSCs. Using this combination1, hESCs or hiPSCs rapidly differentiated into neurons with more than
75% efficiency within 10 days of differentiation. The resulting neurons present classical markers and functional properties
of human nociceptors, including tetrodotoxin (TTX)-resistant, SCN10A-dependent sodium currents and response to
nociceptive stimuli such as ATP and capsaicin1.
This differentiation process is rapid, efficient and can be easily scaled-up. Modulating cell fate of hPSC by Repro-nLS3i or
similar strategies provides a method for basic stem cell research and drug discovery.
Details:
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Chemical Formulation: |
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Repro-nLS (K6570-500) (day 0-5) |
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1000 x DMSO stock |
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0.1 mM of LDN193189 (BMP inhibitor), and |
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solution |
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10 mM of SB431542 (TGF-ß1 inhibitor). |
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Repro-n3i (K6571-500) (day 2-10) |
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3 mM of CHIR99021 (GSKß inhibitor), |
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10 mM of DAPT (Notch inhibitor), and |
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10 mM of SU5402 (FGFR inhibitor) |
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Purity: |
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>98% (for each compound) |
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Shipping condition: |
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The product is shipped in a glass vial at ambient temperature. |
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Storage: |
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Store DMSO solution at -20oC. |
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Usage: |
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Dilute 1000x Repro-nLS stock solution at 1:1000 into your basic ESC/iPSC media. |
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tApply it to the cells on days 0-5. |
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Dilute 1000x Repro-n3i stock solution in Repro-nLS3i at 1:1000 into your basic |
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ESC/iPSC media. Apply it to the cells on days 2-10. |
References:
1. Chambers SM, et al. Combined small-molecule inhibition accelerates developmental timing and converts human
pluripotent stem cells into nociceptors. Nat Biotechnol. 2012. 30(7):715-20.
2. Chambers SM, et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling.
Nat Biotechnol. 2009 27(3):275-80.
3. Yu PB., et al. BMP type I receptor inhibition reduces heterotopic [corrected] ossification Nat Med. 2008. 14(12):
1363-9.
Product Name: Repro-n3i (1000 x)
Repro-n3i is a cocktail of three small molecule inhibitors. According to literature1-3, the combination of these inhibitors
significantly accelerated the process of deriving postmitotic neurons from human ESCs and iPSCs. When used together
with Repro-nLS1, hESCs or hiPSCs rapidly differentiated into neurons with more than 75% efficiency within 10 days of
differentiation. The resulting neurons present classical markers and functional properties of human nociceptors, including
tetrodotoxin (TTX)-resistant, SCN10A-dependent sodium currents and response to nociceptive stimuli such as ATP and
capsaicin1. This differentiation process is rapid, efficient and can be easily scaled-up.
Details:
|
Chemical Formulation: |
: |
3 mM of CHIR99021 (GSKß inhibitor), |
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1000 x DMSO stock |
|
10 mM of DAPT (Notch inhibitor), and |
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solution |
: |
10 mM of SU5402 (FGFR inhibitor) |
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|
|
|
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Purity: |
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>98% (for each compound) |
|
Shipping condition: |
|
The product is shipped in a glass vial at ambient temperature. |
|
Storage: |
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Store DMSO solution at -20oC. |
|
Usage: |
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Dilute 1000x Repro-n3i stock solution at 1:1000 into your basic ESC/iPSC media to |
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reconstitute 1xRepro-n3i media. |
References:
1. Chambers SM, et al. Combined small-molecule inhibition accelerates developmental timing and converts human
pluripotent stem cells into nociceptors. Nat Biotechnol. 2012. 30(7):715-20.
2. Chambers SM, et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling.
Nat Biotechnol. 2009 27(3):275-80.
3. Yu PB., et al. BMP type I receptor inhibition reduces heterotopic [corrected] ossification Nat Med. 2008. 14(12):
1363-9.
Product Name: Repro-nLS (1000 x)
To achieve the full potential of stem cells in regenerative medicine and disease modeling, high conversion efficiency from
hESCs/hiPSCs to neural cells is crucial. A supplement of this small molecule cocktail in cell culture media simplifies the
process and increases the productivity of neural and neuronal cell generation.
Repro-nLS (also named as LSB in reference 1) is a combination of two small-molecule inhibitors (LDN193189 and
SB431542). Under the Repro-nLS condition, both BMP (ALK2/3) and TGF-beta 1(ALK4/5/7) signaling pathways are
inhibited by LDN193189 and SB431542 respectively. hESCs/hiPSCs can be efficiently converted into neural precursor
cells in a monolayer fashion under this dual SMAD inhibition. More than 80% of hESCs are rapidly and uniformly converted into Pax6+ neural precursor cells. This cocktail can be further combined with other patterning factors to readily generate
subtype specific neural and neuronal cells 1-3, such as nociceptors when used with 3i 1 (SU5402, CHIR99021 and DAPT)
(See Repro-nLSB3i).
Details:
|
Chemical Formulation: |
: |
0.1 mM of LDN193189 (BMP inhibitor), and |
|
1000 x DMSO stock |
|
10 mM of SB431542 (TGF-ß1 inhibitor). |
|
solution |
: |
|
|
Purity: |
|
>98% (for each compound) |
|
Shipping condition: |
|
The product is shipped in a glass vial at ambient temperature. |
|
Storage: |
|
Store DMSO solution at -20oC. |
|
Usage: |
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Add/Dilute Repro-nLS stock solution at 1:1000 into your hESC/iPSC basal media to |
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|
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reconstitute 1x media for neural differentiation. |
References:
1. Chambers SM, et al., Combined small-molecule inhibition accelerates developmental timing and converts human
pluripotent stem cells into nociceptors. Nat Biotechnol. 2012 30(7):715-20.
2. Chambers SM, et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling.
Nat Biotechnol. 2009 27(3):275-80.
3. Yu PB., et al. BMP type I receptor inhibition reduces heterotopic ossification. Nat Med. 2008 14(12):1363-9.
Product Name: Repro-nLSC (1000 x)
Data has shown direct transdifferentiation between different germ layers when transfected with a cocktail of defined
transcription factor genes1,3. A supplement of this small molecule cocktail in cell culture media simplifies the process and
increases the productivity of neural and neuronal cell generation.
Repro-nLSC is a cocktail of three small molecule inhibitors. It includes LDN193189, SB431542 and CHIR99021. Using the
combination of synergistic SMAD inhibition through BMP and TGF-beta pathways together with specific GSK-3beta
inhibition, a recent study has presented that postnatal human fibroblasts (HPF), transduced with Ascl1 and Ngn2,
efficiently converted into functional neuron-like cells with yields up to >200% and neuronal purities up to >80%. In
addition to the conversion of mesenchymal cells to induced neurons (iNs), this method has also been tested in the
conversion of other somatic cell types (eg. Cord blood-derived stem cells) to iNs3.
Details:
|
Chemical Formulation: |
: |
0.5 mM of LDN193189 (BMP inhibitor), |
|
1000 x DMSO stock |
|
10 mM of SB431542 (TGF-ß1 inhibitor), and |
|
solution |
: |
2 mM of CHIR99021 (GSK3ß inhibitor) |
|
|
|
|
|
Purity: |
|
>98% (for each compound) |
|
Shipping condition: |
|
The product is shipped in a glass vial at ambient temperature. |
|
Storage: |
|
Store DMSO solution at -20oC. |
|
Usage: |
|
Dilute Repro-nLSC stock solution (1000x) at 1:1000 into your reprogramming media to |
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initiate neuronal conversion. |
References:
1. Vierbuchen T, et al.Direct conversion of fibroblasts to functional neurons by defined factors. Nature. 2010. 463
(7284):1035-41.
2. Chambers SM, et al. Highly efficient neural conversion of human ES and iPS cells by dual inhibition of SMAD signaling.
Nat Biotechnol. 2009 27(3):275-80.
3. Ladewig J., et al. Small molecules enable highly efficient neuronal conversion of human fibroblasts. Nat Methods. 2012.
9(6):575-8.
Ordering information
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Catalog No. |
Product Name |
Size |
|
K7228-500 |
Repro-PACT™ |
0.5mL of 1000 x
0.2mL of 5000 x
1.0mL of 5000 x |
|
K8447-500 |
Repro-ThiPSC |
0.5mL of 1000 x |
|
K6720-500 |
Repro-nSC (1000 x) |
0.5mL of 1000 x |
|
K6572-500 |
Repro-nLS3i (1000 x) |
0.5mL of 1000 x |
|
K6571-500 |
Repro-n3i (1000 x) |
0.5mL of 1000 x |
|
K6570-500 |
Repro-nLS (1000 x) |
0.5mL of 1000 x |
|
K5722-500 |
Repro-nLSC (1000 x) |
0.5mL of 1000 x |
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