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A rapid method for obtaining murine bone marrow cells in high yield
M Levite1, A Meshorer, Y Reisner
1Department of Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Abstract:
A rapid and efficient method for obtaining murine bone marrow cells is described, which yields up to twice the amount of cells obtained by the conventional method of flushing through the bones. The femoral and tibial bones are partially broken by an Omni-Mixer in the presence of phosphate-buffered saline to allow their bone marrow content to extrude into the liquid suspension. Murine bone marrow cells obtained by this method were found to be more than 95% viable, and their differential counts were comparable to those of bone marrow suspensions obtained by the flushing method. Moreover, no contamination by cells from the bone or other surrounding tissues has been observed. Transplantation of bone marrow, obtained by the new extrusion method and depleted of T cells, resulted in long-term stable chimeras in which the hematopoietic reconstitution was comparable to that found in mice transplanted with bone marrow obtained by the flushing method. This new method for obtaining murine bone marrow cells may serve as a time- and mice-sparing alternative to the conventional flushing method, and may also prove useful in other animal models.
Insights
A new method efficiently extracts more murine bone marrow cells by breaking bones, offering a faster, viable alternative to traditional flushing for research and transplantation.
Area of Science:
- * Hematology
- * Immunology
- * Cell Biology
Background:
- * Conventional murine bone marrow extraction relies on flushing, which can be time-consuming and yield limited cell quantities.
- * Optimizing bone marrow cell yield and viability is crucial for preclinical research and transplantation studies.
- * Existing methods may lead to contamination from surrounding tissues.
Purpose of the Study:
- * To introduce a novel, rapid, and efficient method for isolating murine bone marrow cells.
- * To compare the yield, viability, and purity of bone marrow cells obtained by the new method versus the conventional flushing technique.
- * To evaluate the functional capacity of bone marrow cells obtained via the new method in transplantation models.
Main Methods:
- * Murine femoral and tibial bones were partially fractured using an Omni-Mixer in phosphate-buffered saline.
- * Bone marrow cells were extruded into the suspension, followed by collection and processing.
- * Cell viability and differential counts were assessed, and T-cell-depleted bone marrow was used for transplantation studies.
Main Results:
- * The extrusion method yielded up to twice the number of bone marrow cells compared to flushing.
- * Over 95% cell viability was achieved with the new method, with comparable differential counts to the flushing method.
- * Transplantation of extruded bone marrow cells resulted in stable chimeras with effective hematopoietic reconstitution, and no observed contamination.
Conclusions:
- * The described extrusion method provides a significantly more efficient and time-sparing alternative for obtaining murine bone marrow cells.
- * This technique ensures high cell viability and purity, making it suitable for transplantation and other research applications.
- * The method's efficacy and potential applicability in other animal models warrant further investigation.