CC131
Żel ECM zakwalifikowany do komórek macierzystych
Stem cell qualified basement membrane extract (BME) for human ES and iPS cell culture.
Synonim(y):
Ekstrakt z błony podstawnej (BME)
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| Rozmiar/SKU | Dostępność | Cena netto |
|---|---|---|
5 mL | Sprawdź dostępność w koszyku | 2200,00 zł |
5 mL | Skontaktuj się z Obsługą Klienta, aby uzyskać informacje na temat dostępności | 19 790,00 zł |
Informacje o tej pozycji
eCl@ss:
32160801
UNSPSC Code:
12352207
NACRES:
NA.71
Pomoc techniczna
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Pozwól nam pomócform
gel form
technique(s)
cell culture | stem cell: suitable
General description
Stem Cell Qualified ECM Gel to rozpuszczalny ekstrakt błony podstawnej (BME) oczyszczony z guza Engelbreth-Holm-Swarm (EHS), który został wstępnie zakwalifikowany do hodowli ludzkich komórek ES/iPS. Matryca polimeryzuje w temperaturze 20-40°C, tworząc odtworzoną błonę podstawną. ECM Gel zawiera między innymi lamininę, kolagen IV, entaktynę i siarczan heparanu. Matryca została dokładnie przetestowana i zoptymalizowana pod kątem hodowli ludzkich komórek ES/iPS.
Cechy i korzyści
- Eliminuje potrzebę czasochłonnych badań wstępnych i identyfikacji partii.
- Zapewnia zoptymalizowaną powłokę ECM niezbędną do długotrwałej hodowli pluripotencjalnych ludzkich komórek ES/iPS.
- Umożliwia hodowlę ludzkich komórek ES/iPS bez podajnika w pożywce niezawierającej surowicy.
- Kompatybilny ze wszystkimi bezkarmnikowymi pożywkami do hodowli ludzkich komórek ES/iPS (np. mTeSR®, PluriSTEM).
Cechy i korzyści
- Eliminuje potrzebę czasochłonnych badań wstępnych i identyfikacji partii.
- Zapewnia zoptymalizowaną powłokę ECM niezbędną do długotrwałej hodowli pluripotencjalnych ludzkich komórek ES/iPS.
- Umożliwia hodowlę ludzkich komórek ES/iPS bez podajnika w pożywce niezawierającej surowicy.
- Kompatybilny ze wszystkimi bezkarmnikowymi pożywkami do hodowli ludzkich komórek ES/iPS (np. mTeSR®, PluriSTEM).
Stem Cell Qualified ECM Gel Coating Protocol
NOTE: Expansion of human pluripotent stem cells requires culture ware that is coated with prequalified ECM proteins. Below are general guidelines for the coating of 6-well plates using the Stem Cell Qualified ECM Gel (CC131). All procedure should be performed under aseptic conditions in a biological safety cabinet.
1. Thaw the Stem Cell Qualified ECM Gel in the 2-8°C fridge one day before use. Once thawed, maintain ECM Gel on ice at all times and use pre-cooled medium and pipettes to avoid gelling of the product.
IMPORTANT: Do not thaw ECM Gel at temperatures higher than 15ºC to avoid gelling.
2. Dilute the ECM Gel 1:80 with cold DMEM/F12 (D6421) medium. Scale up or down according to the volumes required. Below is an example for coating a 6-well plate with 1.5 mL of 1:80 diluted ECM Gel.
• To make a 1:20 dilution, add 0.5 mL ECM Gel to 9.5 mL cold DMEM/F12 (D6421) or DMEM (D5796) medium into a 15 mL conical tube. Total volume = 10 mL.
• A further 4-fold dilution is required to make the final 1:80 dilution. Add 2.5 mL of 1:20 diluted ECM Gel to 7.5 mL cold DMEM/F12 medium. Total volume = 10 mL
NOTE: The recommended dilution is 1:80, however more concentrated Stem Cell Qualified ECM Gel may be used if desired.
3. Add 1.5 mL of the 1:80 diluted Stem Cell Qualified ECM Gel to each well of a 6-well plate. Swirl the culture plates to spread the ECM Gel evenly across the surface of the plate. Store in a 2 -8°C fridge overnight or at least 2 hours in the fridge before use. If not used immediately, parafilm wrap the ECM coated culture plates and store at 2-8°C until ready to use. Use the ECM coated culture plates within 3-4 days.
4. Prior to seeding the cells, bring the plate back to room temperature for 10-15 minutes, remove the coating solution and add 3 mL/well of human ES/iPSC growth media. Cells can now be plated onto the newly coated plates.
IMPORTANT: Do not allow the plates to dry out.
Frequently Asked Questions (FAQs)
1. I have leftover Stem Cell Qualified ECM Gel (1:20 or 1:80 dilution). How long can I keep it?
Diluted ECM Gel can be kept in the fridge for 3-4 days. Within this time frame, you can either use directly for coating or further dilute to make up the 1:80 dilution. Avoid multiple freeze thaw cycles.
2. Can precoated plates be frozen for future use?
No, we do not recommend freezing pre-coated plates. Pre-coated plates should be sealed with Parafilm and may be stored at 2-8°C for up to 3-4 days before use.
3. Can I use a dilution factor higher than 1:80?
We recommend a maximum dilution of 1:80. Further optimization may be required for higher dilutions.
4. My Stem Cell Qualified ECM Gel has already polymerized? How do I make it liquid again?
Once polymerized, ECM Gel cannot revert to a liquid form again. It is recommended to start with a fresh frozen aliquot and coat the plates again. To prevent polymerization, store ECM Gel on ice during handling.
5. Do I need to filter the Stem Cell Qualified ECM Gel before use?
No, ECM Gel is cell culture tested and ready-to-use once diluted.
NOTE: Expansion of human pluripotent stem cells requires culture ware that is coated with prequalified ECM proteins. Below are general guidelines for the coating of 6-well plates using the Stem Cell Qualified ECM Gel (CC131). All procedure should be performed under aseptic conditions in a biological safety cabinet.
1. Thaw the Stem Cell Qualified ECM Gel in the 2-8°C fridge one day before use. Once thawed, maintain ECM Gel on ice at all times and use pre-cooled medium and pipettes to avoid gelling of the product.
IMPORTANT: Do not thaw ECM Gel at temperatures higher than 15ºC to avoid gelling.
2. Dilute the ECM Gel 1:80 with cold DMEM/F12 (D6421) medium. Scale up or down according to the volumes required. Below is an example for coating a 6-well plate with 1.5 mL of 1:80 diluted ECM Gel.
• To make a 1:20 dilution, add 0.5 mL ECM Gel to 9.5 mL cold DMEM/F12 (D6421) or DMEM (D5796) medium into a 15 mL conical tube. Total volume = 10 mL.
• A further 4-fold dilution is required to make the final 1:80 dilution. Add 2.5 mL of 1:20 diluted ECM Gel to 7.5 mL cold DMEM/F12 medium. Total volume = 10 mL
NOTE: The recommended dilution is 1:80, however more concentrated Stem Cell Qualified ECM Gel may be used if desired.
3. Add 1.5 mL of the 1:80 diluted Stem Cell Qualified ECM Gel to each well of a 6-well plate. Swirl the culture plates to spread the ECM Gel evenly across the surface of the plate. Store in a 2 -8°C fridge overnight or at least 2 hours in the fridge before use. If not used immediately, parafilm wrap the ECM coated culture plates and store at 2-8°C until ready to use. Use the ECM coated culture plates within 3-4 days.
4. Prior to seeding the cells, bring the plate back to room temperature for 10-15 minutes, remove the coating solution and add 3 mL/well of human ES/iPSC growth media. Cells can now be plated onto the newly coated plates.
IMPORTANT: Do not allow the plates to dry out.
Frequently Asked Questions (FAQs)
1. I have leftover Stem Cell Qualified ECM Gel (1:20 or 1:80 dilution). How long can I keep it?
Diluted ECM Gel can be kept in the fridge for 3-4 days. Within this time frame, you can either use directly for coating or further dilute to make up the 1:80 dilution. Avoid multiple freeze thaw cycles.
2. Can precoated plates be frozen for future use?
No, we do not recommend freezing pre-coated plates. Pre-coated plates should be sealed with Parafilm and may be stored at 2-8°C for up to 3-4 days before use.
3. Can I use a dilution factor higher than 1:80?
We recommend a maximum dilution of 1:80. Further optimization may be required for higher dilutions.
4. My Stem Cell Qualified ECM Gel has already polymerized? How do I make it liquid again?
Once polymerized, ECM Gel cannot revert to a liquid form again. It is recommended to start with a fresh frozen aliquot and coat the plates again. To prevent polymerization, store ECM Gel on ice during handling.
5. Do I need to filter the Stem Cell Qualified ECM Gel before use?
No, ECM Gel is cell culture tested and ready-to-use once diluted.
Application
Research Category
Stem Cell Research
Stem Cell Research
Preparation Note
Przechowywać w temperaturze -20°C. Rozmrażać przez noc na lodzie w lodówce w temperaturze 2-8°C. Zminimalizować ryzyko rozmrożenia poprzez podzielenie na podwielokrotności jednorazowego użytku. Przechowywać podwielokrotności w zamrażarce o temperaturze -20°C lub -80°C, aż będą gotowe do użycia. Nie przechowywać w zamrażarce zabezpieczonej przed mrozem.
Uwaga: Stem Cell Qualified ECM Gel jest niezwykle wrażliwy na temperaturę i może polimeryzować w temperaturze pokojowej lub wyższej w ciągu 5-10 minut. Przechowywać przez cały czas w lodzie i używać wstępnie schłodzonych końcówek pipet, probówek i płytek.
Uwaga: Stem Cell Qualified ECM Gel jest niezwykle wrażliwy na temperaturę i może polimeryzować w temperaturze pokojowej lub wyższej w ciągu 5-10 minut. Przechowywać przez cały czas w lodzie i używać wstępnie schłodzonych końcówek pipet, probówek i płytek.
Analysis Note
Appearance: Red, clear of particulates
Endotoxin Level: <20 EU/mL
Microbial Contamination: Pass
Mycoplasma: Negative
Functional Assay: <5% spontaneous differentiation for 3 passages using Human iPS cells
Endotoxin Level: <20 EU/mL
Microbial Contamination: Pass
Mycoplasma: Negative
Functional Assay: <5% spontaneous differentiation for 3 passages using Human iPS cells
Legal Information
mTeSR is a registered trademark of WiCell Research Institute, Inc.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
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Ta pozycja | |||
|---|---|---|---|
| description Stem cell qualified basement membrane extract (BME) for human ES and iPS cell culture. | description growth-factor reduced, without phenol red, liquid, BioReagent, suitable for cell culture | description liquid, BioReagent, suitable for cell culture | description - |
| form gel form | form liquid | form liquid | form - |
| technique(s) cell culture | stem cell: suitable | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | mammalian: suitable |
Klasa składowania
12 - Non Combustible Liquids
wgk
WGK 1
flash_point_f
Not applicable
flash_point_c
Not applicable
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