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A Bone of Contention: Best Approach for Harvesting Bone Marrow to Maximize TNC and CD34 Cell Counts

Introduction. Bone marrow (BM) has been utilized as a source of stem cells for transplantation for many years. Although the use of BM has decreased with the advent of mobilized stem cells, utilization is increasing once again due to the lower rate of chronic GVHD associated with BM as a stem cell source. There is no generally accepted technique for harvesting BM. Protocols vary both in relation to the volume of each aspirate and the number of aspirates performed at each puncture site. .

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A Bone of Contention: Best Approach for Harvesting Bone Marrow to Maximize TNC and CD34 Cell Counts

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    1. A Bone of Contention: Best Approach for Harvesting Bone Marrow to Maximize TNC and CD34+ Cell Counts

    3. Aim To investigate whether differences in the volume and number of aspirates taken at the time of BM harvest affect the number of mononuclear cells (particularly CD34+ cells and CD3+ cells) collected.

    4. Method Consented adult donors were positioned in the prone position for collection of bone marrow from the posterior iliac crests. Triplicate samples of 5, 10 and 20ml aspirate volumes were taken at the start. Subsequent harvesting was performed using 10ml aspirate volumes. Triplicate samples were taken from the 10ml aspirates when harvest volumes of 250, 500, 750 and 1000mls were attained. Samples were analysed for total nucleated cell (TNC) count, CD34+ and CD3+ cell counts. See Figure 1

    6. Results

    9. Figure 4. Cell numbers obtained from 10ml marrow aspirates at various times during marrow harvest.

    11. Conclusion The aspiration of increasing volumes of BM does not produce a proportional increase in total numbers of TNC, CD34+ or CD3+ cells collected. The highest ratio of CD34+ to CD3+ cells is observed with 10ml aspirates. There is a progressive drop in yield of both CD34+ and CD3+ cells once 750ml of BM has been aspirated in 10 ml aliquots. Continued harvesting after collection of 1000ml will produce small numbers of CD34+ cells. If insufficient stem cells have been obtained at this time, a second harvest may be a better option than continued aspiration on the same occasion.

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