Immortalized MPC’s using Er-HoxB8
Faculty Mentor
Jason Ashley
Presentation Type
Poster
Start Date
4-14-2026 11:30 AM
End Date
4-14-2026 1:30 PM
Location
PUB NCR
Primary Discipline of Presentation
Biology
Abstract
Many biological experiments rely on isolation of primary cells from mouse tissues to investigate cellular and physiological processes. Experiments using murine bone marrow naturally yield low densities of primary macrophages, creating during primary cell culture. These low densities require additional mouse sacrifices for multiple experiments. To reduce animal use and generate a renewable macrophage progenitor source, experiments are being performed using the ER-HoxB8 conditional immortalization system to establish a myeloid progenitor cell line. ER-HoxB8 is an estrogen regulated transcription factor system that, in the presence of β-estradiol, blocks cell differentiation and maintains progenitors in a proliferating state. Granulocyte Macrophage Colony-Stimulating Factor (GM-CSF) is also added during to maintain myeloid progenitors prior to differentiation into macrophages after release from HoxB8 mediated arrest. Removal of β-estradiol along with the addition of Macrophage Colony-Stimulating Factor (M-CSF) drives myeloid progenitors into macrophages. To create the immortalized cell line, progenitors were first isolated from murine bone marrow by flushing hind-leg bones. After cytokine pre-stimulation of murine bone marrow progenitor cells, ER-HoxB8 carried in a MSCV retroviral plasmid was transfected into Platinum-E packaging cells to produce ecotropic retrovirus and then transduced. Cells derived from the immortalized line successfully differentiated into osteoclasts and have been used in other experiments.
Recommended Citation
VerStrate, Camden, "Immortalized MPC’s using Er-HoxB8" (2026). 2026 Symposium. 27.
https://dc.ewu.edu/srcw_2026/ps_2026/p2_2026/27
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Immortalized MPC’s using Er-HoxB8
PUB NCR
Many biological experiments rely on isolation of primary cells from mouse tissues to investigate cellular and physiological processes. Experiments using murine bone marrow naturally yield low densities of primary macrophages, creating during primary cell culture. These low densities require additional mouse sacrifices for multiple experiments. To reduce animal use and generate a renewable macrophage progenitor source, experiments are being performed using the ER-HoxB8 conditional immortalization system to establish a myeloid progenitor cell line. ER-HoxB8 is an estrogen regulated transcription factor system that, in the presence of β-estradiol, blocks cell differentiation and maintains progenitors in a proliferating state. Granulocyte Macrophage Colony-Stimulating Factor (GM-CSF) is also added during to maintain myeloid progenitors prior to differentiation into macrophages after release from HoxB8 mediated arrest. Removal of β-estradiol along with the addition of Macrophage Colony-Stimulating Factor (M-CSF) drives myeloid progenitors into macrophages. To create the immortalized cell line, progenitors were first isolated from murine bone marrow by flushing hind-leg bones. After cytokine pre-stimulation of murine bone marrow progenitor cells, ER-HoxB8 carried in a MSCV retroviral plasmid was transfected into Platinum-E packaging cells to produce ecotropic retrovirus and then transduced. Cells derived from the immortalized line successfully differentiated into osteoclasts and have been used in other experiments.
