Department of Transfusion Medicine, Clinical Center, National Institutes of Health, Building 10, Room 1C711, Bethesda, MD 20892-1184, USA.
J Transl Med. 2012 Feb 6;10:23. doi: 10.1186/1479-5876-10-23.
Bone marrow stromal cells (BMSCs) are being used to treat a variety of conditions. For many applications a supply of cryopreserved products that can be used for acute therapy is needed. The establishment of a bank of BMSC products from healthy third party donors is described.
The recruitment of healthy subjects willing to donate marrow for BMSC production and the Good Manufacturing Practices (GMP) used for assessing potential donors, collecting marrow, culturing BMSCs and BMSC cryopreservation are described.
Seventeen subjects were enrolled in our marrow collection protocol for BMSC production. Six of the 17 subjects were found to be ineligible during the donor screening process and one became ill and their donation was cancelled. Approximately 12 ml of marrow was aspirated from one posterior iliac crest of 10 donors; one donor donated twice. The BMSCs were initially cultured in T-75 flasks and then expanded for three passages in multilayer cell factories. The final BMSC product was packaged into units of 100 × 106 viable cells, cryopreserved and stored in a vapor phase liquid nitrogen tank under continuous monitoring. BMSC products meeting all lot release criteria were obtained from 8 of the 11 marrow collections. The rate of growth of the primary cultures was similar for all products except those generated from the two oldest donors. One lot did not meet the criteria for final release; its CD34 antigen expression was greater than the cut off set at 5%. The mean number of BMSC units obtained from each donor was 17 and ranged from 3 to 40.
The production of large numbers of BMSCs from bone marrow aspirates of healthy donors is feasible, but is limited by the high number of donors that did not meet eligibility criteria and products that did not meet lot release criteria.
骨髓基质细胞（BMSCs）被用于治疗多种疾病。对于许多应用，需要提供可用于急性治疗的冷冻保存产品。本文描述了从健康的第三方供体中建立 BMSC 产品库的情况。
描述了招募愿意捐献骨髓用于 BMSC 生产的健康受试者，以及用于评估潜在供体、采集骨髓、培养 BMSCs 和 BMSC 冷冻保存的良好生产规范（GMP）。
17 名受试者参加了我们的骨髓采集协议，用于 BMSC 生产。在供体筛选过程中，有 6 名受试者不符合条件，1 名受试者患病，其捐献被取消。从 10 名供体的一个后髂嵴中抽吸了约 12ml 骨髓；一名供体捐献了两次。BMSCs 最初在 T-75 培养瓶中培养，然后在多层细胞工厂中扩增三代。最终的 BMSC 产品被包装成 100×106 个活细胞的单位，冷冻保存并在连续监测下储存在气相液氮罐中。从 11 次骨髓采集中有 8 次获得了符合所有批次放行标准的 BMSC 产品。除了来自两个最年长供体的产品外，所有产品的原代培养物的生长速度相似。有一个批次不符合最终放行标准；其 CD34 抗原表达大于设定的 5%的截止值。从每个供体获得的 BMSC 单位数量平均为 17 个，范围从 3 到 40 个。
从健康供体的骨髓抽吸物中大量生产 BMSC 是可行的，但受到许多不符合资格标准的供体和不符合批次放行标准的产品的限制。
