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The aim of the study was to investigate whether normal aging leads to blood cells being produced from fewer cell clones. This was examined by comparing which X chromosome the cells were using between different age groups and between neutrophils and T cells.
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In other words, they studied human hematopoiesis across age by analyzing purified blood cell lineages from the subjects.
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Modell/material
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- 197 hematologically normal females
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- 23 new born
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@@ -6,23 +10,12 @@ Modell/material
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- purified neutrofils
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- purified T-cells
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The model is _human hematopoiesis across age_ and the material is _lineage-purified blood cells_
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methods:
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1. cell separation (neutrofil and T-cell)
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2. x-chromosome inactivation analysis (humana PCR)
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3. T-cell clonality check
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> [!NOTE]
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> They separated neutrophils and T cells, measured X-inactivation skewing, and checked that T cells were not dominating.
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### **What was compared / tested**
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- **Age groups:** newborn vs young adults vs elderly
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- **Cell lineages:** neutrophils vs T cells
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- **Readout:** degree of X-chromosome inactivation skewing
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**Methods**
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1. Peripheral blood samples were collected
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2. Neutrophils and T cells were isolated using cell separation techniques _(centrifugation followed by lineage-specific purification)_
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3. X-chromosome inactivation was analyzed using the HUMARA PCR assay _(PCR-based measurement of which X chromosome is active)_
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4. T cells were analyzed by PCR to check for clonal expansion _(T-cell receptor γ gene rearrangement analysis)_
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### Results
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1. X chromosome usage is similar in newborns and young adults.
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2. Neutrophils from elderly individuals show strongly uneven X chromosome usage.
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