Human CD34+ HSPCs were cultured as previously described.5 (link),20 (link) Human CB-CD34+ HSPCs were generously provided by the Binns Program for Cord Blood Research at Stanford University. Plerixafor mPB-CD34+ HSPCs were purchased from AllCells (Alameda, CA, USA) or received from the Stanford LCGM Facility. The cells were thawed per the manufacturer’s instructions and were cultured at 37°C, 5% CO2, and 5% O2 for 48 h (48-h pre-stimulation) or 72 h (72-h pre-stimulation) prior to genome editing. Cell pre-stimulation medium and culturing medium were the same, which was GMP SCGM medium from CellGenix (Portsmouth, NH, USA) supplemented with human cytokine cocktail (PeproTech, Rocky Hill, NJ, USA) containing stem cell factor (SCF) (100 ng/mL), thrombopoietin (100 ng/mL), Fms-like tyrosine kinase 3 ligand (100 ng/mL), and interleukin 6 (100 ng/mL). UM171 (35 nM) (StemCell Technologies, Vancouver, BC, Canada), streptomycin (20 mg/mL), and penicillin (20 U/mL) were added into the cell culture medium.
Gmp scgm medium
Manufactured by CellGenix
Sourced in United States
GMP SCGM medium is a cell culture medium designed for the expansion and maintenance of human mesenchymal stem cells (hMSCs) and other cell types in a Good Manufacturing Practice (GMP) environment. The medium provides the necessary nutrients and growth factors to support the growth and proliferation of cells while adhering to GMP standards.
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Lab products found in correlation
2 protocols using gmp scgm medium
1
Optimized ex vivo expansion of HSPCs
2
Expansion of CD34+ HSPCs from Mobilized Peripheral Blood and Cord Blood
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Frozen CD34+ HSPCs derived from mobilized peripheral blood (mPB) were purchased from AllCells and thawed according to manufacturer’s instructions. CD34+ HSPCs from cord blood were either purchased frozen from AllCells or acquired from donors under informed consent via the Binns Program for Cord Blood Research at Stanford University and used fresh without freezing51 (link). The works was performed on blinded samples from healthy donors of CD34+ HSPCs and inclusive of all cells that were donated to research. The research was performed, analyzed and communicated in an ethical manner.
Cord blood-derived CD34+ cells were used for all in vitro experiments. unless noted otherwise. mPB derived CD34+ HSPCs were used for all transplantation experiments shown in Fig.5 and in vitro experiments shown in Fig. 2c–e . All CD34+ HSPCs were cultured in GMP SCGM medium (Cellgenix) supplemented with SCF (100 ngml–1), TPO (100 ngml–1), Flt3 ligand (100 ngml–1), IL-6 (100 ngml–1), and StemRegenin1 (0.75 mM). Cells were cultured at 37 °C, 5% CO2, and 5% O2.
Cord blood-derived CD34+ cells were used for all in vitro experiments. unless noted otherwise. mPB derived CD34+ HSPCs were used for all transplantation experiments shown in Fig.
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