UCB mononuclear cells were separated by density gradient centrifugation with lymphoprep (Boster Bio, China), and then CD34+ HSCs were purified from the mononuclear cell fraction by using a human CD34-positive selection kit (Stem Cell Technology, Vancouver, Canada) according to the manufacturer’s protocol. For expansion of CD34+ cells, hematopoietic growth factors were added as follows: IL-3 10 ng/mL, SCF 100 ng/mL, TPO 100 ng/mL, Flt-3 ligand 100 ng/mL (PeproTech, USA). About 4×104 cells were seeded in each plate well. The cells were cultured at 37°C in a humidified atmosphere of 5% CO2 and harvested on Day 0, 6, and 12. Cell pellets and supernatants were stored at −80°C for further study.
Lymphoprep
Manufactured by Boster Bio
Sourced in China
Lymphoprep is a density gradient medium used for the isolation of mononuclear cells from whole blood or bone marrow. It facilitates the separation of mononuclear cells by centrifugation.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using lymphoprep
1
CD34+ HSCs Expansion and Culture
2
Isolation of CD34+ Hematopoietic Cells
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Bone marrow cell suspensions (10 mL) were obtained, respectively, by bone marrow paracentesis from the patients and healthy donors, for isolation of CD34+ hematopoietic cells. Mononuclear cells were then obtained by density gradient centrifugation with Lymphoprep (Boster Bio, China). CD34+ hematopoietic cells were isolated by anti-CD34 (BD Biosciences, Franklin, NJ, USA) antibodies and the MACS system (Stem Cell Technology, Vancouver, Canada) from the mononuclear cells according to the manufacturer's protocols.
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