Silvestri F, Banavali S, Yin M, Gopal V, Savignano C, Baccarani M, Preisler H D
University of Cincinnati Medical Center, OH.
Haematologica. 1992 Jul-Aug;77(4):307-10.
Pluripotent hemopoietic stem cells, progenitors of all hemolymphopoietic lineages, and clonogenic cells from many patients with acute nonlymphocytic leukemia (ANLL) and chronic myeloid leukemia (CML) express the CD34 antigen on their surface. Isolation of these cell populations is of primary experimental and clinical importance.
Six bone marrow (BM) and 10 peripheral blood (PB) samples were obtained from 2 normal individuals, 3 patients with CML and 9 with ANLL. The CD34+ cell fraction was isolated using MY10 antibody, sheep anti-mouse immunomagnetic beads and the enzyme chymopapain. Indirect immunofluorescence and semisolid culture were employed to evaluate the percentage of CD34+ cells and that of clonogenic cells in each cell fraction.
The frequency of CD34+ cells in the original unseparated populations was (mean +/- SE) 24.3 +/- 7.3%, and reached 85.0 +/- 2.7% in the isolated CD34-positive fractions; in the negative fractions it was only 2.7 +/- 1.7%. According to these results, the great majority of clonogenic cells was separated in the CD34-positive fractions and depleted in those CD34-negative. Moreover, chymopapain was shown to be non-toxic to the clonogenic cells.
Positive immunoselection using My10 Ab, immunomagnetic beads and chymopapain is a method for isolating almost pure progenitors from the BM and PB of normal individuals and patients with myeloid leukemias.
多能造血干细胞是所有造血淋巴系的祖细胞，许多急性非淋巴细胞白血病（ANLL）和慢性粒细胞白血病（CML）患者的克隆形成细胞在其表面表达CD34抗原。分离这些细胞群体具有重要的实验和临床意义。
从2名正常个体、3名CML患者和9名ANLL患者中获取6份骨髓（BM）和10份外周血（PB）样本。使用MY10抗体、羊抗鼠免疫磁珠和木瓜凝乳蛋白酶分离CD34+细胞组分。采用间接免疫荧光和半固体培养法评估各细胞组分中CD34+细胞和克隆形成细胞的百分比。
原始未分离群体中CD34+细胞的频率为（均值±标准误）24.3±7.3%，在分离的CD34阳性组分中达到85.0±2.7%；在阴性组分中仅为2.7±1.7%。根据这些结果，绝大多数克隆形成细胞在CD34阳性组分中被分离出来，而在CD34阴性组分中减少。此外，木瓜凝乳蛋白酶对克隆形成细胞无毒。
使用My10抗体、免疫磁珠和木瓜凝乳蛋白酶进行阳性免疫选择是一种从正常个体和髓系白血病患者的骨髓和外周血中分离几乎纯的祖细胞的方法。
