Sorted human CXCL12hi/LEPR+ cells were seeded at a density of 1·5 × 102 cells/cm2 into 96‐well plates and were cultured in MSC expansion medium for 10 days. Then medium was removed, and 5 000 human cord blood CD34+ cells (RIKEN BRC, Tsukuba, Japan) were co‐cultured with or without the adherent CXCL12hi/LEPR+ cells in StemSpan Serum‐Free Expansion Medium (StemCell Technologies, Vancouver, BC, Canada) supplemented with 25 ng/ml of recombinant human SCF (Fuji Film Wako Pure Chemical), thrombopoietin (TPO; Fuji Film Wako Pure Chemical) and fms‐related receptor tyrosine kinase 3 ligand (FLT3L; Fuji Film Wako Pure Chemical), and 1% penicillin/streptomycin at 37°C with 5% CO2. After one week of co‐culture, cells were harvested and the numbers of CD34+ cells were counted using flow cytometry.
Thrombopoietin tpo
Manufactured by Fujifilm
Sourced in Canada, United States
Thrombopoietin (TPO) is a glycoprotein that functions as a cytokine and a growth factor. It is primarily responsible for the regulation of platelet production and maturation. TPO is a core component in various laboratory and research applications.
Automatically generated - may contain errors
Lab products found in correlation
Flt3l, by Fujifilm (1 mentions)
Stemspan serum free expansion medium, by STEMCELL (1 mentions)
Millicell cell culture plates, by Merck Group (1 mentions)
Flt 3 ligand, by Fujifilm (1 mentions)
Stemspan, by STEMCELL (1 mentions)
Vascular endothelial growth factor (vegf), by R&D Systems (1 mentions)
Penicillin streptomycin, by Welgene (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
2 protocols using thrombopoietin tpo
1
Expansion of Human Hematopoietic Stem Cells
2
Coculture Analysis of CD34+ Cells
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Coculture analysis was performed in 12-well Millicell Cell Culture Plates (0.4 µm pore size; Millipore, Billerica, MA, USA) using Stem Span serum-free medium (StemCell Technologies), supplemented with 50 ng/mL VEGF, 20 ng/mL IL-6 (R&D Systems), 100 ng/mL SCF (Kirin), 20 ng/mL thrombopoietin (TPO; Wako), 100 ng/mL Flt-3 ligand (Wako), and 1% penicillin-streptomycin (WELGENE). Human CD34+ cells isolated from HUCB were seeded in the lower compartment of the transwell, and the transwell membrane inserts were seeded with human CD34− cells isolated from HUCB or left unseeded. To measure the EPC-CFU potential of CD34+ cells after coculture, the transwell inserts were removed after 7 d. The expansion of CD34+ cell was determined by cell counting.
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