Lundell B I, Mandalam R K, Smith A K
Aastrom Biosciences, Inc., Ann Arbor, MI 48105, USA.
J Hematother. 1999 Apr;8(2):115-27. doi: 10.1089/106161299320398.
Ex vivo expansion of bone marrow (BM) mononuclear cells (MNC) in a perfused culture system produces stem-progenitor cell type(s) in sufficient number(s) for hematopoietic reconstitution. The limitations in using fresh BM MNC for ex vivo expansion include additional cell processing and inflexibility in patient treatment. Cryopreservation of whole bone marrow (WBM) eliminates processing costs of MNC or CD34+ cell selection and allows for flexibility in patient treatment. We developed a convenient system to cryopreserve and thaw small volume WBM aspirations (n = 13) and then compared the expandability of unprocessed normal cryopreserved/thawed (C/T) WBM to that of fresh BM MNC cultured in the presence of erythropoietin, PIXY 321, and Flt3-ligand. Ex vivo expansion potential was retained in WBM aspirates after C/T. When initiated with 225 million viable nucleated cells, clinical scale expansion cultures (n = 6) yielded 9.7+/-2.8 x 10(8) total cells, which contained 10.4+/-5.8 x 10(6) colony-forming units-granulocyte-macrophage (CFU-GM), 1.3+/-1.4 x 10(4) LTCIC, and 2.2 x 10(6) CD34+Lin- cells, sufficient cell numbers for clinical use. These studies demonstrate that ex vivo perfusion culture expansion of unfractionated C/T WBM (< or =30 ml) provides doses of stem-progenitor cells similar in composition to expanded fresh BM MNC, previously demonstrated to achieve hematopoietic reconstitution.
在灌注培养系统中对骨髓（BM）单个核细胞（MNC）进行体外扩增可产生数量充足的用于造血重建的干祖细胞类型。使用新鲜BM MNC进行体外扩增的局限性包括额外的细胞处理以及患者治疗缺乏灵活性。全骨髓（WBM）的冷冻保存消除了MNC或CD34 +细胞选择的处理成本，并为患者治疗提供了灵活性。我们开发了一种便捷的系统来冷冻保存和解冻小体积的WBM抽吸物（n = 13），然后将未处理的正常冷冻保存/解冻（C/T）WBM与在促红细胞生成素、PIXY 321和Flt3配体存在下培养的新鲜BM MNC的可扩展性进行比较。C/T后WBM抽吸物保留了体外扩增潜力。当以2.25亿个有活力的有核细胞开始时，临床规模的扩增培养（n = 6）产生了9.7±2.8×10⁸个总细胞，其中包含1.04±0.58×10⁶个集落形成单位 - 粒细胞 - 巨噬细胞（CFU - GM）、1.3±1.4×10⁴个长期培养起始细胞（LTCIC）和2.2×10⁶个CD34⁺Lin⁻细胞，这些细胞数量足以用于临床。这些研究表明，未分级的C/T WBM（≤30 ml）的体外灌注培养扩增可提供与扩增的新鲜BM MNC组成相似的干祖细胞剂量，先前已证明其可实现造血重建。
