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. In other words, they studied human hematopoiesis across age by analyzing purified blood cell lineages from the subjects. ----- ## Study - 1 97 hematologically normal females - 23 newborn - 94 young adult (median 30) - 80 eldery (over 75) - purified neutrofils - purified T-cells ---- ### Metoder 1. Peripheral blood samples were collected 2. Neutrophils and T cells were isolated using cell separation techniques _(centrifugation followed by lineage-specific purification)_ 3. X-chromosome inactivation was analyzed using the HUMARA PCR assay _(PCR-based measurement of which X chromosome is active)_ 4. T cells were analyzed by PCR to check for clonal expansion _(T-cell receptor γ gene rearrangement analysis)_ --- ### Results 1. X chromosome usage is similar in newborns and young adults. 2. Neutrophils from elderly female individuals show strongly uneven X chromosome usage. 3. T-cells show much less uneven X chromosome usage and no dominant clonal expansion --- ### Cord blood neutrophils ![[image-183.png|148x156]] - Graph shows the distribution of X chromosome usage in 23 newborn - X-axis: % of cells using the less common X chromosome. - Y-axis: number of individuals in each group. Conclusion: Newborns mostly show balanced use of the two X chromosomes --- ### > 75 years neutrophils ![[image-184.png|145x158]] - This graph shows the purified neutrofils from 80 elderly (y >= 75) females - X and Y axis is the same as in previous graph Conclusion: Elderly neutrophils frequently show strongly uneven X chromosome usage. ---- ### Questions - What was the main question the authors wanted to answer? - Why did they choose this specific method instead of another one? - What does this figure show in one sentence? - What is the main conclusion from your article? - What would be the biggest limitation of this study?