Cell line/type | iPSc (induced Pluripotent Stem Cells) |
---|---|
Species | Human |
Animal free | Yes |
Product | E8 medium Chou et al. 2015 |
Chou, B. K., Gu, H., Gao, Y., Dowey, S. N., Wang, Y., Shi, J., ... & Cheng, L. (2015). A Facile Method to Establish Human Induced Pluripotent Stem Cells From Adult Blood Cells Under Feeder‐Free and Xeno‐Free Culture Conditions: A Clinically Compliant Approach. Stem cells translational medicine, 4(4), 320-332. In this study, highly efficient iPS cell derivation under feeder-free and xeno-free conditions is reported via three major improvements. First, a combination of three EBNA1/OriP episomal vectors was revised expressing five transgenes, which increased reprogramming efficiency by >/=10-50-fold from our previous vectors. Second, human recombinant vitronectin proteins were used as cell culture substrates, alleviating the need for feeder cells or animal-sourced proteins. Finally,the previously critical step of manually picking individual iPS cell clones by pooling newly emerged iPS cell colonies was eliminated. Pooled cultures were then purified based on the presence of the TRA-1-60 pluripotency surface antigen, resulting in the ability to rapidly expand iPS cells for subsequent applications. These new improvements permit a consistent and reliable method to generate human iPS cells with minimal clonal variations from blood MNCs, including previously difficult samples such as those from patients with paroxysmal nocturnal hemoglobinuria. Mononuclear cells were cultured in IMDM/F12 supplemented with different components as specified in the Materials and Methods, under the heading; Isolation and Culture of PB MNCs Under a Serum-Free Condition (expanding mononuclear cells). Human iPS cells were cultured in E8 medium as specified in the Materials and Methods, under the heading; Expansion of Human iPS Cells in the E8 Medium on Vitronectin as Adhesion Substrates. The formulation of E8 medium and the protocol for its preparation is listed in a previous article, Sharma et al. 2015. Sharma, A., Li, G., Rajarajan, K., Hamaguchi, R., Burridge, P. W., & Wu, S. M. (2015). Derivation of highly purified cardiomyocytes from human induced pluripotent stem cells using small molecule-modulated differentiation and subsequent glucose starvation. Journal of visualized experiments: JoVE, (97). https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4367508/pdf/sctm_20140214.pdf |
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Source | Literature - own formulation |
Chemically defined > Yes | Yes |
Contains phenol red > No | No |
Antibiotics free > Yes | Yes |