iPSC culture kit (Qk512)

Protein background

Human induced pluripotent stem cells (iPSC) are an in vitro model that are functionally immortal and have the capacity to proliferate while maintaining the ability to, hypothetically, differentiate to any somatic cell type. Interest in iPSC generation has grown significantly because they can be used across various research areas and fields without facing the same ethical concerns associated with the use of embryonic stem cells.

This has created a need to culture large quantities of individual iPSC lines or a diverse array of different iPSC lines. Consequently, this has increased the cost and time-consuming demand for daily media changes. Maintaining pluripotency during routine maintenance is a challenge with cell cultures known for their sensitivity, such as iPSC. Using high purity and thermostable growth factors can help reduce this risk as they reduce the impact on genotypic stability as they lack impurities such as endotoxins and provide a more consistent exposure to necessary growth factors.

Vitronectin is a multifunctional glycoprotein commonly used extracellular matrix for the culture of iPSCs on. This is due to its key roles in supporting cell adhesion, proliferation, and differentiation. By interacting with integrins on the cell surface, vitronectin creates a favorable environment for iPSC to attach and spread, ensuring robust and stable cell adhesion. Vitronectin helps in maintaining the pluripotent state of iPSC by interactions with signaling pathways that support the expression of key pluripotency markers and genes, provides survival signals that help reduce cell apoptosis and is known to provide extracellular matrix cues that mimic the natural cellular environment to support differentiate into specific cell types.

E8-like media is a chemically defined media, meaning that it contains less undefined components than are found in other complex media. This results in reduced variability and improves the reproducibility of the culture between an individual iPSC line or a variety of different iPSC lines. This formulation has been optimized to support robust cell proliferation while ensuring the pluripotency of iPSC remains and resulting in improved cell yields and more consistent scaling up of cultures. Pluripotency of iPSCs can be evaluated by investigating intracellular surface markers such as NANOG, SOX2 and OCT-4.