Sharp T G, Sachs D H, Matthews J G, Maples J, Woody J N, Rosenberg S A
J Immunol Methods. 1984 Apr 27;69(2):187-95. doi: 10.1016/0022-1759(84)90317-x.
Harvest of human bone marrow directly from freshly resected bone provides purer preparations of marrow than can be obtained by the conventional technique of multiple aspirations from the iliac crests. In particular, directly harvested marrow is much less heavily contaminated with peripheral blood lymphocytes, a known source of mature T cells. Because of the possible relevance of these contaminating T cells for cadaveric bone marrow transplants, the best source of human marrow harvested directly from bone has been studied. Human bone marrow was harvested from 46 surgical specimens and 9 cadaveric tissue donors. Vertebral bodies provided the best source of bone marrow with average yields of 3.1 +/- 1.6 X 10(9) cells per vertebra. When entire ilia were removed and processed for marrow, an average of 1.6 +/- 1.0 X 10(9) cells was obtained. Surgically resected ribs yielded lower amounts of marrow with a mean cell number of 3.2 +/- 2.6 X 10(8) per rib. Isolation of bone marrow mononuclear cells from these preparations by density gradient centrifugation resulted in a loss of 45% of the starting cells. Human bone marrow was found to contain 5-6% T cells before gradient separation and these cells were immunologically competent as measured in vitro by responses to mitogens and alloantigens. This technique may be useful in obtaining human bone marrow for both immunologic studies and allogeneic transplantation.
直接从新鲜切除的骨中采集人骨髓，比通过传统的从髂嵴多次抽吸技术获得的骨髓制剂更纯净。特别是，直接采集的骨髓受外周血淋巴细胞污染的程度要低得多，外周血淋巴细胞是成熟T细胞的已知来源。由于这些污染的T细胞可能与尸体骨髓移植有关，因此对直接从骨中采集的人骨髓的最佳来源进行了研究。从46个手术标本和9个尸体组织供体中采集了人骨髓。椎体是骨髓的最佳来源，每个椎体的平均产量为3.1±1.6×10⁹个细胞。当整个髂骨被切除并处理以获取骨髓时，平均获得1.6±1.0×10⁹个细胞。手术切除的肋骨产生的骨髓量较低，每根肋骨的平均细胞数为3.2±2.6×10⁸个。通过密度梯度离心从这些制剂中分离骨髓单个核细胞导致起始细胞损失45%。在梯度分离前发现人骨髓含有5%-6%的T细胞，这些细胞在体外通过对有丝分裂原和同种异体抗原的反应来测量具有免疫活性。这项技术可能有助于获得用于免疫研究和同种异体移植的人骨髓。
