Ivanovic Zoran, Duchez Pascale, Morgan Doris A, Hermitte Francis, Lafarge Xavier, Chevaleyre Jean, Praloran Vincent, Dazey Bernard, Vezon Gérard, Boiron Jean-Michel
French Blood Establishment Aquitaine-Limousin, Bordeaux, France.
Transfusion. 2006 Jan;46(1):118-25. doi: 10.1111/j.1537-2995.2005.00677.x.
Used leuko-depletion filters (LDFs), containing billions of white blood cells (WBCs), are discarded. Because the steady-state blood contains low quantities of stem and progenitor cells that are retained in LDFs, the viability and the functional properties of mononuclear cells (MNCs) and CD 34+ cells recovered from LDFs were investigated.
WBCs were recovered from LDFs by use of a closed system. MNCs and CD 34+ cells were isolated from freshly LDF-recovered WBCs or after their overnight incubation. The CD 34+ cells were enumerated, as well as the number of colony-forming unit (CFU)-granulocyte-macrophage, burst-forming unit-erythroid, and CFU-Mixed. The expansion in clinical-scale volume cultures (serum-free medium plus stem cell factor, granulocyte-colony-stimulating factor, and megakaryocyte growth and development factor) was performed starting from MNCs, freshly isolated CD 34+ cells, and CD 34+ cells isolated after overnight incubation of WBCs. The erythroid, megakaryocytic, eosinophilic, and monocyte-myelocytic lineage differentiation of LDF-recovered CD 34+ cells was challenged in liquid cultures by adding relevant cytokines.
Nearly 450 x 10(3) viable CD 34+ cells were recovered per LDF. These cells exhibit unimpaired colony-forming ability. It is possible to expand these cells ex vivo, but their response to cytokines is different compared to mobilized peripheral blood and cord blood CD 34+ cells. Thus, further work is necessary to optimize their ex vivo expansion. These cells give rise to the mature cells and precursors of erythroid, megakaryocytic, eosinophilic, and monomyelocytic lineage in liquid cultures.
MNCs and CD 34+ cells recovered from the LDFs exhibit unimpaired functional capacities. Recent development of ex vivo technologies for expansion, retro-differentiation, and differentiation reinforces the value in cell therapy of these LDG-recovered peripheral blood progenitor cells that are routinely discarded.
使用过的白细胞滤除器（LDFs）含有数十亿白细胞（WBCs），通常被丢弃。由于稳态血液中含有少量保留在LDFs中的干细胞和祖细胞，因此对从LDFs中回收的单核细胞（MNCs）和CD 34+细胞的活力和功能特性进行了研究。
通过封闭系统从LDFs中回收WBCs。从新鲜回收的LDF-WBCs或过夜培养后分离出MNCs和CD 34+细胞。对CD 34+细胞进行计数，并统计集落形成单位（CFU）-粒细胞-巨噬细胞、爆式红系集落形成单位和CFU-混合集落的数量。从MNCs、新鲜分离的CD 34+细胞以及WBCs过夜培养后分离的CD 34+细胞开始，在临床规模的体积培养物（无血清培养基加干细胞因子、粒细胞集落刺激因子和巨核细胞生长发育因子）中进行扩增。通过添加相关细胞因子，在液体培养中挑战LDF回收的CD 34+细胞的红系、巨核系、嗜酸性和单核-髓系谱系分化。
每个LDF可回收近450×10³个有活力的CD 34+细胞。这些细胞表现出未受损的集落形成能力。有可能在体外扩增这些细胞，但与动员的外周血和脐血CD 34+细胞相比，它们对细胞因子的反应不同。因此，有必要进一步开展工作以优化其体外扩增。这些细胞在液体培养中产生红系、巨核系、嗜酸性和单核-髓系谱系的成熟细胞和前体细胞。
从LDFs中回收的MNCs和CD 34+细胞表现出未受损的功能能力。体外扩增、逆向分化和分化技术的最新进展强化了这些常规被丢弃的LDF回收外周血祖细胞在细胞治疗中的价值。
