Project description:To investigate the impact of radiation damage on hematopoietic stem cell fate decision, we conducted a longitudinal study, collecting single-cell data on days 2 post-radiation exposure. This approach enabled us to observe changes in the subpopulations of hematopoietic stem cells at the LSK level in mice over time. Additionally, we assessed alterations in their fate determination and lineage differentiation tendencies throughout the recovery process.
Project description:In this study, we aimed to investigate the differentiation dynamics of LSK (Lin-Sca1+c-Kit+) cells following radiation injury. LSK cells, which are a critical component of the hematopoietic stem and progenitor cell population, play a vital role in the regeneration and repair of the hematopoietic system after damage. By utilizing these cells, we sought to understand the mechanisms underlying their response to radiation, focusing on their capacity for self-renewal and differentiation into various blood cell lineages.
Project description:To investigate the impact of perturbing key differentiation driver genes on hematopoietic stem cell fate decision, we employed a Dapp1 gene knockout approach. This allowed us to observe changes in the subpopulations of hematopoietic stem cells at the LSK level in mice. Additionally, we assessed alterations in their fate decision and lineage differentiation tendencies
Project description:Gene expression of LSK (lin-Sca-Kit+) hematopoietic stem cells from wild type mice was compared with LSK from Cebpa knock-in mutant mice (K/K, K/L, and L/L mutants). Fetal liver cells for each genotype were competitively transplanted into irradiant recipients. Donor-derived LSK cells were isolated by FACS sorting of recipient bone marrow. 3 biological replicates of each were generated and expression profiles were determined by hybridization to Affymetrix Moe430_2 arrays.
Project description:To further identify gene expression signatures in the niche cells (CD45 negative) during proliferation of Hematopoietic stem cells (LSK), we employed mice whole genome (60K) microarray expression profiling as a discovery platform to identify the up-regulated and down-regulated genes of the niche. [Samples A-D] In this experiment a physiological stress model was created where the recipient mice were subjected to sub lethal radiation (700 CGy) following a transplantation of 30,000 LSK cells (HSCs). Bone marrow cells were isolated on day 0 (before transplantation) and day 10 (post transplantation of 30,000 LSK cells when maximum proliferation of HSCs was observed). Donor HSCs was sorted by FACS following RNA isolation and cDNA synthesis followed by single color global gene expression analysis. Agilent one-color experiment,Organism:Mouse, Agilent Whole Genome Mouse 8x60k (AMADID: 26986) , Labeling kit: Agilent Quick-Amp labeling Kit (p/n5190-0442)
Project description:Analysis of hematopoietic LSK(Lin-Sca1+c-Kit+) cells lacking the Serum response factor (SRF) gene. Results provide insight into the role of SRF in regulating genetic programs important for hematopoietic stem cell development
Project description:Little is known of hematopoietic stem (HSC) and progenitor (HPC) cell self-renewal. The role of Brahma (BRM), a chromatin remodeler, in HSC/HPC function is unknown. Bone marrow (BM) from Brm-/- mice manifested increased numbers of long- and short-term HSCs, GMPs, and increased numbers and cycling of functional HPCs. Brm-/- HSC/HPC show demonstrated functional differences compared to wildtype HSC/HPC in vivo and ex vivo, due in part to increased intracellular valine in lineage negative BM. To determine changes in gene programs associated with loss of BRM, we performed RNA sequencing on Brm-/- LSK cells. Brm-/- LSK exhibited upregulated interferon response/cell cycle gene programs, suggesting important extrinsic effects on primitive HSC/HPC from more mature HPC due to the absence of BRM.