stemdiff tm apel 2 medium  (STEMCELL Technologies Inc)

Journal: Stem Cells and Development

Article Title: Enhanced Ex Vivo Generation of Erythroid Cells from Human Induced Pluripotent Stem Cells in a Simplified Cell Culture System with Low Cytokine Support

doi: 10.1089/scd.2019.0132

Figure Lengend Snippet: Schematic illustration of the cell culture system. (1) Hematopoietic induction: colonies of undifferentiated hiPSCs were transferred into low-binding plates to induce EB formation. After 5 days, spherical EBs were further cultured on adherent plates in APEL ™ medium containing SCF, EPO, and IL-3. The medium was changed weekly. Within 2 weeks, an HCFC was established, from which hematopoietic cells were continuously released into the supernatant and harvested for further characterization. (2) Erythroid differentiation: cells released into the supernatant were harvested and differentiated into RBCs in a three-phase erythropoiesis system over 18 days. Hematopoietic and erythroid differentiation were monitored by colony formation assay, flow cytometry, microscopy, and hemoglobin analysis. EB, embryoid body; EPO, erythropoietin; HCFC, hematopoietic cell forming complex; hiPSCs, human induced pluripotent stem cells; IL-3, interleukin 3; RBCs, red blood cells; SCF, stem cell factor.

Article Snippet: Thereafter, 10–15 spherical EBs were transferred to six-well tissue culture plates (Sarstedt) with STEMdiff TM APEL TM 2 Medium (STEMCELL Technologies), 5% PFHM-II Protein-Free Hybridoma Medium (Thermo Fisher Scientific), 5 ng/mL interleukin 3 (IL-3; PeproTech), 100 ng/mL stem cell factor (SCF; PeproTech), and 3 U/mL erythropoietin (EPO; Erypo; Janssen Biologics B.V.).

Techniques: Cell Culture, Binding Assay, Colony Assay, Flow Cytometry, Microscopy