Project description:LLC1 cells were injected into the tibia of DMP1-Cre PhAM mice to study the intercellular mitochondrial transfer in the bone microenvironment. We used single-cell RNA sequencing (scRNA-seq) to analyze the cellular heterogeneity of osteocyte-derived mitochondria recipient cells.
Project description:We injected LLC1 cells into tibia to construct a murine model to study bone microenvironment in cancer bone metastasis. We used single cell RNA sequencing (scRNA-seq) to analyze the cellular heterogeneity of tumor-infiltrating cells.
Project description:Primary human skeletal muscle cells (Lonza) were treated with LLC1 conditioned medium, LLC1 conditioned medium plus Calcitriol, LLC1 non-conditioned medium or LLC1 non-conditioned medium plus Calcitriol for a period of 24 hours prior to isolation of RNA.
Project description:Bone regeneration remains clinically challenging, partly due to limited knowledge of the cellular mechanisms governing bone repair. Osteocytes, which constitute the majority of bone cells, form an dendritic network and are central regulators of skeletal homeostasis. Residing in a mineralized and low-oxygen environment, they are exposed to hypoxic conditions that may shape their function, yet the role of hypoxia in osteocyte behavior and angiogenic signaling during bone regeneration is still unclear. This study investigated the effects of hypoxia on osteocytes and their paracrine interaction with endothelial cells. MLO-Y4 osteocytic cells were cultured under normoxia (21% O₂), moderate hypoxia (5% O₂), or severe hypoxia (1% O₂), and conditioned media were applied to mouse endothelial cells. Transcriptomic analyses were performed following optimization of the co-culture model. Hypoxia promoted osteocyte dendritic network development and induced oxygen-dependent metabolic and transcriptional adaptations. Severe hypoxia induces a stronger hypoxic response and glycolytic shift compared with moderate hypoxia, leading to distinct osteocyte profiles. In contrast, osteocyte-conditioned media did not significantly affect endothelial gene expression, suggesting that hypoxia-independent mechanisms may govern osteocyte-endothelial communication. These findings highlight oxygen availability as a key regulator of osteocyte physiology and underscore the need for further investigation into its role during bone regeneration.
Project description:CTCF ChIP-seq on osteocyte originated from osteocyte For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODE_Data_Use_Policy_for_External_Users_03-07-14.pdf
Project description:Control ChIP-seq on osteocyte originated from osteocyte For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODE_Data_Use_Policy_for_External_Users_03-07-14.pdf
Project description:Type of Experiment: 1) Profiles of Osteoblast vs. osteocyte in vitro; 2) Profiles of osteoblast low density vs. confluency in vitro; 3) Profiles of osteocyte with gap junction vs. without gap junction. Experimental factors: 1) 2T3 osteoblast cells at low density expressed extensive filopodia, reminiscent of early osteoblast precursors and similar to MLO-Y4 dendritic processes. 2) MLO-Y4 osteocytes at low vs. high density represent the genes that are changed in a highly connected network vs. low connected network. The number of hybridizations performed: Triplicate hybridizations for each status. Keywords = Osteoblast Keywords = Osteocyte