Chemically defined conditions for human iPSC derivation and culture

Abstract

We reexamine the individual components for human ES and iPS cell culture, and formulate a cell culture system in which all protein reagents for liquid media, attachment surfaces, and splitting are chemically defined. A major improvement is the lack of a serum albumin component, as variations in either animal or human sourced albumin batches have previously plagued human ES and iPS cell culture with inconsistencies. Using this new medium (E8) and vitronectin-coated surfaces, we demonstrate improved derivation efficiencies of vector-free human iPS cells with an episomal approach. This simplified E8 medium should facilitate both the research use and clinical applications of human ES and iPS cells and their derivatives, and should be applicable to other reprogramming methods.

High-efficiency motor neuron differentiation from human pluripotent stem cells and the function of Islet-1

“…As a first step towards attaining this goal, we made several modifications to the neural induction procedure. First, we cultured hESCs in the recently developed E8 medium 15 and on ECM substrates consisting of fibronectin, vitronectin and laminin, before compound C induction (Fig. 1a,c).…”

“…H1 and H9 cells were cultured on irradiated mouse embryonic fibroblasts in media containing DMEM/F12, 20% knockout serum replacement, 4 ng ml À 1 basic fibroblast growth factor (Invitrogen), 1% non-essential amino acid, 1 mM glutamine and 0.1 mM b-mercaptoethanol. For feeder-independent conditions, hESCs were cultured on Matrigel (BD Biosciences)-coated plates in mTeSR1 medium (StemCell Technologies) or E8 medium for the support of long-term self-renewal 15,43 . The quality and high viability of the cells are essential for their proper responses to induction of differentiation.…”

Animal‐cell culture media: History, characteristics, and current issues

“…These cells initially were cultivated on a layer of feeder cells in a serum‐ or KSR‐supplemented medium,104 but in anticipation of clinical applications, the efforts shifted to the development of culture conditions that are feeder‐free and xeno‐free (Table 5). 105, 106, 107, 108, 109, 110, 111 Among these, the E8 medium that was developed by Guokai Chen et al. has become popular.…”

Promise and challenges of human iPSC-based hematologic disease modeling and treatment

“…Besides the virus-free integration-free reprogramming methods, conditions for mouse feeder-free iPSC culture are being constantly improved to make iPSCs safer and more suitable for clinical applications [37][38][39][40][41]. However, it is still a challenge to establish systems that can sufficiently support human iPSC self-renewal, genetic modifications and differentiation while compatible with clinical production under current Good Manufacturing Practice (cGMP) standards.…”