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Updated: Mar 19, 2026
Flow Cytometry Analysis of Murine Bone Marrow Hematopoietic Stem and Progenitor Cells and Stromal Niche Cells
Published on: September 28, 2022
Vortex-dislodged cells from bone marrow trephine biopsy yield satisfactory results for flow cytometric
K Bommannan1, M U S Sachdeva2, M Gupta1
1Department of Hematology, Postgraduate Institute of Medical Education & Research, Chandigarh, India.
Insights
Flow cytometric immunophenotyping (FCI) is feasible using cells from trephine biopsy (TB) specimens. This method provides rapid hematologic diagnosis, especially in difficult bone marrow aspiration cases.
Area of Science:
- Hematology
- Oncology
- Cellular Biology
Background:
- Accurate bone marrow (BM) sampling is critical for diagnosing hematologic disorders.
- Unsuccessful bone marrow aspiration (BMA) hinders flow cytometric immunophenotyping (FCI) in hematologic malignancies.
- Alternative diagnostic methods often provide less information than FCI.
Purpose of the Study:
- To assess the feasibility of FCI using cells obtained from trephine biopsy (TB) specimens.
- To establish an alternative method for FCI when BMA is unsuccessful.
Main Methods:
- Trephine biopsy (TB) cores were collected from patients with pancytopenia or dry tap BMA.
- Cores were vortexed in phosphate-buffered saline to dislodge cells.
- The resulting cell pellet was used for flow cytometric immunophenotyping (FCI).
Main Results:
- 34 dry tap cases were analyzed.
- Cell pellets suitable for FCI were obtained in 94% (32/34) of cases via vortexing.
- FCI provided a rapid diagnosis in 100% (32/32) of cases with available cell pellets.
Conclusions:
- Vortex-dislodged cells from TB specimens are a viable alternative for FCI.
- This method improves diagnostic yield in challenging bone marrow cases.
- The procedure is efficient, particularly for resource-limited settings, reducing repeat BMA procedures.
Introduction:
A good bone marrow (BM) sample is essential in evaluating many hematologic disorders. An unsuccessful BM aspiration (BMA) procedure precludes a successful flow cytometric immunophenotyping (FCI) in most hematologic malignancies. Apart from FCI, most ancillary diagnostic techniques in hematology are less informative. We describe the feasibility of FCI in vortex-dislodged cell preparation obtained from unfixed trephine biopsy (TB) specimens.
Methods:
In pancytopenic patients and dry tap cases, routine diagnostic BMA and TB samples were complemented by additional trephine biopsies. These supplementary cores were immediately transferred into sterile tubes filled with phosphate-buffered saline, vortexed, and centrifuged. The cell pellet obtained was used for flow cytometric immunophenotyping.
Results:
Of 7955 BMAs performed in 42 months, 34 dry tap cases were eligible for the study. Vortexing rendered a cell pellet in 94% of the cases (32 of 34), and FCI rendered a rapid diagnosis in 100% of the cases (32 of 32) where cell pellets were available.
Conclusion:
We describe an efficient procedure which could be effectively utilized in resource-limited centers and reduce the frequency of repeat BMA procedures.
