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The Gibco Essential 8 Adaptation Kit includes Gibco Essential 8 Medium and recombinant human rhLaminin-521 (rhLaminin-521) for optimal survival during transition from a feeder-dependent to feeder-free culture system.
rhLaminin-521 has been proven to promote cellular survival under stressful conditions in the absence of small molecule inhibitors (Figure 1). In addition, rhLaminin-521 has been shown to support PSC growth for >20 passages without any signs of karyotypic abnormalities and to maintain the ability of PSCs to differentiate into all three germ line lineages (Figure 2). When used in conjunction with Essential 8 Medium, rhLaminin-521 enables a trouble-free transition from feeder-based media to a xeno-free, feeder-free media system.
Figure 1. Optimum recovery of PSCs during transition to Gibco Essential 8 Medium in the absence of small molecule inhibitors. An episomal CD34+-derived iPSC line was cultured on mouse irradiated feeders in KSR-based medium prior to transition to Essential 8 Medium. For transition, colonies were passaged using collagenase IV per the protocol provided with the Essential 8 Adaptation Kit and seeded into Essential 8 Medium in wells coated with rhLaminin-521 (green diamonds), Gibco Geltrex matrix (blue circles), or rhVTN-N (red triangles). Confluency of cultures was monitored using the IncuCyte ZOOM™ system post-passage (n=3 per condition).
Figure 2. Human ESCs maintain expression of self-renewal factors. Human embryonic stem cells (ESCs) were grown under feeder-free conditions using Gibco Essential 8 Medium for 20 passages in wells coated with rhLaminin-521. The cells were stained for pluripotency markers using the PSC 4-Marker ICC Kit.
The following components are recommended to create a complete media system for routine expansion and maintenance of PSCs with Essential 8 Medium.
For Research Use Only. Not for use in diagnostic procedures.