Gibco™ RevitaCell™ Supplement (100X)

Catalog No.	Quantity
A2644501	5 mL

Catalog No. A2644501 Supplier Gibco™ Supplier No. A2644501

Description

RevitaCell™ Supplement (100X)

RevitaCell™ Supplement (100X) has been optimized for use with pluripotent stem cells (PSCs), either as a post-thaw recovery solution to improve cell viability or in combination with Essential 8™ Medium to enhance single cell passaging applications. It can also be used for recovery of primary neurons, traditionally a challenging cell to efficiently recover. The significant improvement seen in these applications is due to the presence of a more specific ROCK inhibitor coupled with molecules containing antioxidant and free radical scavenger properties that minimize the impact of stress on the cells.

Enabling—provides maximum cell viability when performing single cell passaging

Rapid cell recovery—high post-thaw cell viability and growth of pluripotent stem cells (passaged as clumps or single cells) and neurons

Consistent—formulated using animal origin-free and chemically defined materials for quality and confidence

Single cell passaging
Use of the RevitaCellTM Supplement for 18-24 hours post-single cell passaging provides vast improvement in cell survival. Cells are shown to reach passaging confluency within 3-4 days post-seeding.

Higher specificity
Kinase profiling of pro-survival small molecules in RevitaCell™ Supplement shows higher specificity in comparison to traditional ROCK inhibitors (i.e., Y-27632 and Thiazovivin).

Quicker post-thaw recovery
PSCs recovered at 40,000 viable cells/cm² using media supplemented with RevitaCell™ Supplement for the first 24 hours post-thaw recovery followed by feeding with unsupplemented Essential 8™ Medium will typically reach 80% confluency in <5 days, significantly reducing the average post-thaw recovery time of most cryopreservation solutions. Neurite outgrowth of neurons is also significantly improved using RevitaCell™ Supplement.

Specifications

• Content And Storage: Store at -5 to -20°C and protect from light.
• Cell Type: Stem Cells (iPS - Induced Pluripotent Stem)
• Classification: Animal Origin-free
• Concentration: 100 X
• Culture Type: Stem Cell (Embryonic, iPS - Induced Pluripotent Stem)
• Product Type: Media Supplement
• Manufacturing Quality: cGMP for medical devices, 21 CFR Part 820 and ISO 13485
• Product Line: Gibco, RevitaCell
• Quantity: 5 mL
• Shipping Condition: Dry Ice
• Source: Animal origin-free

Frequently Asked Questions (FAQs)

Can RevitaCell Supplement be used in addition to a traditional ROCK Inhibitor?

No. Combination of RevitaCell Supplement with a traditional ROCK inhibitor will result in deleterious effects on PSCs.

The RevitaCell Supplement in the PSC Cryopreservation Kit has a strong smell. Is this normal?

Yes, it is normal that the RevitaCell Supplement has a strong smell.

What is the difference between the PSC Cryopreservation Kit and the CTS PSC Cyropreservation Kit?

The CTS PSC Cryopreservation Kit (Cat. No. A4239301) provides the same efficient recovery, consistency, and reduced variability as the PSC Cryopreservation Kit (Cat. No. A2644601). However, these two kits have different intended use statements, i.e., “For Research Use or Manufacturing of Cell, Gene, or Tissue-Based Products. CAUTION: Not intended for direct administration into humans or animals” and “For Research Use Only”, respectively. Furthermore, the CTS PSC Cryopreservation Kit has full USP sterility testing info and a Drug Master file (DMF), whereas the sterility testing of the PSC Cryopreservation is via a modified USP method, and it does not have a DMF.

Is StemFlex Medium compatible with downstream differentiation kits available from Thermo Fisher Scientific?

Yes. We have seen compatibility with the following differentiation kits provided by Thermo Fisher Scientific: PSC Cardiomyocyte Differentiation Kit (Cat. No. A2921201), PSC Definitive Endoderm Induction Kit (Cat. No. A3062601), PSC Neural Induction Medium (Cat. No. A1647801), and PSC Dopaminergic Neuron Differentiation Kit (Cat. No. A3147701).

What protocol is recommended for delivery of Cas9 protein:guide RNA complex to my PSCs cultured in StemFlex Medium?

We recommend use of the Neon Electroporation device for electroporation of PSCs with Cas9 protein:guide RNA complex following the protocol guidance in the following demonstrated protocol (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/PG1584-PJT1313-COL31227-Demonstrated-Protocol-Performing-CRISPR-Cas9-FHR.pdf); see the section entitled “Knockout by electroporation of RNP using the Neon Transfection System.” We have seen that the Neon electroporation protocols 7 and 14 provide optimal indel formation while maintaining cell survival. However, the electroporation conditions may need to be optimized for your pluripotent cell line.

I see in the flexible feeding schedules specified for StemFlex Medium that cells should be fed the day after passaging, however this is incompatible with my current workflow. Have you evaluated skipping the feed the day after passaging?

Yes. The main reason that the workflow states this is to accommodate ROCK inhibitor or RevitaCell Supplement that have been added during passaging. In such circumstances where ROCK inhibitor or RevitaCell Supplement are added, you should feed your cells within 18-24 hours post-passaging. However, if ROCK inhibitor or RevitaCell Supplement are not included at the time of passaging, then you may skip the day post-passaging.

Do I need to use ROCK inhibitor during single-cell passaging in StemFlex Medium?

The use of ROCK inhibitor is not required when culturing in StemFlex Medium on rhLaminin-521. However, supplementation with RevitaCell Supplement can provide additional support to PSCs during stressful transitions such as single-cell passaging.

What is the PSC Cryopreservation Kit?

The PSC cryopreservation kit contains xeno-free PSC Cryopreservation Medium, which is a ready-to-use solution for the cryopreservation of early passage pluripotent stem cells (PSCs), and Gibco Revitacell Supplement (100X), a chemically defined recovery supplement for use in the post-thaw culture medium. When used in combination, these reagents help minimize loss of cell viability, maximize post-thaw recovery, and minimize unwanted differentiation of PSCs. This kit can also be used to cryopreserve and recover peripheral blood mononuclear cells (PBMCs) to improve post-thaw cell viability and recovery.

Can Gibco RevitaCell Supplement help cell recovery after electroporation?

Yes, this has been tested with iPSCs. However, testing on primary cells has not been performed.

Should cells be pre-treated with Gibco RevitaCell Supplement for 24 hours before cryopreservation?

No, such pre-treatment results in deleterious effects on the cells.

Can Gibco RevitaCell Supplement be used in addition to a traditional ROCK Inhibitor?

No. Combination of the Gibco RevitaCell Supplement with a traditional ROCK inhibitor results in deleterious effects on PSCs.

Can Gibco RevitaCell Supplement be used in serum-supplemented media for culturing primary cells?

We have not tested this application.

For feeder-free cultures, which types of feeder-free matrix has Gibco RevitaCell Supplement been shown to be compatible with?

Gibco RevitaCell Supplement been shown to be compatible with Gibco Geltrex matrix, Gibco Vitronectin, LN-521 Stem Cell Matrix, and iMatrix-511.

Can Gibco RevitaCell Supplement be used on iPSCs cultured on feeders or is this recommended only for feeder-free culture of iPSC?

Both types of cultures have been tested.

What species of primary cells has Gibco RevitaCell Supplement been used on?

We have only tested it with human species for the following cell types: corneal epithelial cells, keratinocytes, peripheral blood mononuclear cells, and rat cortical neurons.

What species of PSCs has Gibco RevitaCell Supplement been used on?

We have only tested it with human PSCs.

Can I perform single cell passaging with Gibco Essential 8 Flex Medium?

Yes, Essential 8 Flex Medium is compatible with single cell passaging. Whenever you are performing single cell passaging, ensure that you use Gibco RevitaCell Supplement (Cat. No. A2644501) or another ROCK inhibitor.

Can I use ROCK inhibitors with Gibco Essential 8 Flex Medium?

Yes, Essential 8 Flex Medium has been used with ROCK inhibitors. We suggest using Gibco RevitaCell Supplement (Cat. No. A2644501), which has been specifically designed to minimize the impact of stress on PSCs.

Do I need to add a ROCK inhibitor during passaging when culturing in Gibco Essential 8 Flex Medium?

As with many other PSC media, ROCK inhibitor is not required with Essential 8 Flex Medium when Gibco Versene Solution is used to passage PSCs as clusters. If cells are plated as singlets, we suggest that Gibco RevitaCell Supplement (Cat. No A2644501) or another ROCK inhibitor be used.

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov

For Research Use Only. Not for use in diagnostic procedures.