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Updated: Dec 24, 2025
Pan-myeloid Differentiation of Human Cord Blood Derived CD34+ Hematopoietic Stem and Progenitor Cells
Published on: August 9, 2019
Isolation of non-activated monocytes from human umbilical cord blood
Erik Normann1, Thierry Lacaze-Masmonteil, Bonnie Winkler-Lowen
1Department of Women's and Children's Health, Uppsala University Children's Hospital, Uppsala, Sweden. erik.normann@kbh.uu.se
Problem:
Methods for monocyte purification are common but few work with umbilical cord monocytes that do not activate the cell for subsequent culture analysis.
Methods Of Study:
The collection procedure avoids use of needles and procedures that variably activate blood clotting and uses a purification procedure that involves diluted Ficoll, autologous serum to remove platelets and 42% and 51% Percoll step gradients for the final purification. The resulting monocytes were stimulated with bacterial lipopolysaccharide and formalin-treated bacteria Escherichia coli and group B streptococci (GBS) to secrete TNF-alpha and IL-1beta, measured by ELISA.
Results:
The purification procedure results in non-active but stimulation-competent monocytes with high yields (2.3-9 x 10(7) cells) and purity (from 70% to 98%).
Conclusion:
We describe a procedure that is easy, uses common reagents and provides a uniformly high yield and purity of non-activated fetal monocytes for studies of innate defense responses.
Insights
A new method purifies umbilical cord monocytes without activation, yielding high purity and quantity. These non-activated monocytes are suitable for studying innate immune responses.
Area of Science:
- Immunology
- Cell Biology
- Neonatal Research
Background:
- Monocyte purification is crucial for immunological studies.
- Existing methods often activate umbilical cord monocytes, limiting their use in subsequent analyses.
- There is a need for a gentle purification method for fetal monocytes.
Purpose of the Study:
- To develop and validate a non-activating purification method for umbilical cord monocytes.
- To ensure high yield and purity of monocytes for immunological research.
- To provide a reliable source of non-activated fetal monocytes for innate defense studies.
Main Methods:
- A novel collection procedure avoiding needles and clotting activators.
- Purification using diluted Ficoll, autologous serum, and Percoll density gradients.
- Assessment of monocyte function via stimulation with bacterial components (LPS, E. coli, GBS) and measurement of cytokine secretion (TNF-alpha, IL-1beta) by ELISA.
Main Results:
- The procedure yields 2.3-9 x 10^7 monocytes with 70-98% purity.
- Purified monocytes are non-activated but remain stimulation-competent.
- Successful cytokine secretion (TNF-alpha, IL-1beta) upon stimulation was observed.
Conclusions:
- A simple, effective method for purifying non-activated fetal monocytes has been established.
- The procedure utilizes common reagents and provides consistent high yield and purity.
- This method is ideal for studying fetal innate immune responses.
