Control the level of stress on your cells during single-cell passaging with Gibco Essential 8 Medium and Gibco RevitaCell Supplement. Together, they help minimize the impact of stress from single-cell passaging, enabling maximal cell viability and more efficient recovery of cells.

  • Minimize the impact of stress—RevitaCell Supplement supports cells in stressful applications through a blended formulation of a more specific Rho-associated protein kinase (ROCK) inhibitor coupled with antioxidants and free radical scavengers
  • Non-addictive—shown to have a non-addictive impact on downstream cell utility, unlike other ROCK inhibitors
  • Expanded applications—enables use of Essential 8 Medium in applications where cells need additional support

View application note   View protocol

Support through stressful events including single-cell passaging

RevitaCell Supplement can be used for routine single-cell passaging of PSCs in Essential 8 Medium, providing efficient recovery of cells while retaining normal morphology, pluripotency, and karyotype. Furthermore, the prosurvival small molecule contained within this cocktail is shown to be a more selective ROCK inhibitor, minimizing the off-target pathways affected.

Figure 1. Gibco RevitaCell Supplement shows utility in single-cell passaging using Gibco TrypLE Select Enzyme. iPSCs generated using episomal vectors were passaged using TrypLE Select Enzyme and seeded at a density of 25,000 viable cells/cm2 onto culture plates coated with truncated recombinant human vitronectin, in Gibco Essential 8 Medium with 1X RevitaCell Supplement. Following a 24-hour recovery, iPSCs were fed with Essential 8 Medium alone for the remainder of their time in culture.

Six panel phase-contrast image series showing iPSC recovery from 0 hours to 114 hours following single cell passaging.

Figure 2. Normal morphology is retained following single-cell passaging using Gibco Essential 8 Medium plus Gibco RevitaCell Supplement. iPSCs generated using episomal vectors were passaged using Gibco TrypLE Select Enzyme and seeded at a density of 25,000 viable cells/cm2 onto culture plates coated with truncated recombinant human vitronectin, in Essential 8 Medium with 1X RevitaCell Supplement. Following a 24-hour recovery, iPSCs were fed with Essential 8 Medium alone for the remainder of their time in culture.

Complete media system

The following components are recommended to create a complete media system for weekend-free routine expansion and maintenance of PSCs with Gibco Essential 8 Flex Medium.

Note: For single-cell passaging in Essential 8 medium in the absence of a ROCK inhibitor, rhLaminin-521 matrix can be used with Essential 8 Medium. This protocol would not require addition of RevitaCell Supplement. View this protocol reference

Single-cell passaging with Essential 8 Medium Application Note

Culture medium

Matrix

Supplemental reagent

Passaging reagent