University of Utah Cell Therapy and Regenerative Medicine Facility
University of Utah Cell Therapy and Regenerative Medicine Facility.
Ann Clin Lab Sci. 2020 Nov;50(6):781-789.
Major ABO incompatible hematopoietic progenitors from bone marrow (HPC(M)) donor collections that are destined for clinical transplantation are typically processed to deplete products of red blood cells (RBCs). The purpose of this study was to compare RBC depletion when using the Spectra Optia® relative to a 2-step method involving a COBE2991 instrument to obtain a buffy coat followed by a hydroxyethyl starch (HES) density gradient (COBE+HES) of the buffy coat.
Post-processing recoveries of products undergoing 4, 8, and 10 bone marrow processing (BMP) cycles (i.e. 1 cycle=1 volume of HPC(M)) with the Spectra Optia® were determined for volume, RBC content, viable total nucleated cells (vTNC), viable CD34+ cells (vCD34), viable CD3+ cells (vCD3) and colony-forming-cells (CFC). Subsequent RBC depletions with Spectra Optia® were then performed with 10 BMP cycles on additional HPC(M) collections and were compared against a retrospective analysis of historical COBE+HES post-processing data.
Ten BMP cycles of HPC(M) (n=6) products were identified as optimal with volume reductions of 81.3±1.6 % and RBC reductions of 97.0±0.6 % with the Spectra Optia®. This also resulted in an average of 0.28 ±0.14 mL of RBC/kg (mean±SD; n=6) with vTNC yields of 65.0±10.9%, vCD34+ yields of 98.5±12.7%, and vCD3+ yields of 90.6±10.0%. Recoveries with the COBE+HES methodology resulted in vTNC recoveries of 62.9±20.5% (mean±SD; n=30) and 0.63±0.71 mL of RBC/kg (mean±SD; n=30).
The Spectra Optia® is a viable option for depleting HPC(M) harvests of contaminating RBC in situations of ABO incompatibility. Target cells from a MNC rich fractionation were preserved through processing while eliminating RBC contaminants.
从骨髓（HPC(M)）供体采集物中去除主要的 ABO 不相容造血祖细胞，这些祖细胞通常用于临床移植，需要经过处理以去除红细胞（RBC）产物。本研究的目的是比较使用 Spectra Optia®与涉及 COBE2991 仪器获得浓缩白细胞层（CBC），然后用羟乙基淀粉（HES）密度梯度（CBE+HES）分离浓缩白细胞层的两步法（COBE+HES）进行 RBC 去除的效果。
通过 Spectra Optia®对经过 4、8 和 10 个骨髓处理（BMP）循环（即 1 个循环=1 体积 HPC(M)）的产品进行处理后产物的体积、RBC 含量、有活力的总核细胞（vTNC）、有活力的 CD34+细胞（vCD34）、有活力的 CD3+细胞（vCD3）和集落形成细胞（CFC）的回收情况进行了测定。然后，对另外的 HPC(M)采集物进行 10 个 BMP 循环的 Spectra Optia®后续 RBC 去除，并与 COBE+HES 回顾性分析数据进行比较。
确定 10 个 BMP 循环的 HPC(M)（n=6）产品体积减少 81.3±1.6%，RBC 减少 97.0±0.6%，这是使用 Spectra Optia®的最佳条件。这也导致平均每公斤（kg）（n=6）有 0.28±0.14 mL RBC，vTNC 回收率为 65.0±10.9%，vCD34+回收率为 98.5±12.7%，vCD3+回收率为 90.6±10.0%。使用 COBE+HES 方法学，vTNC 回收率为 62.9±20.5%（n=30），每公斤（kg）（n=30）有 0.63±0.71 mL RBC。
在 ABO 不相容的情况下，Spectra Optia®是去除富含 MNC 分离物中污染 RBC 的可行选择。通过处理去除 RBC 污染物，同时保留了来自单核细胞丰富部分的靶细胞。
