Herbein G, Sovalat H, Wunder E, Baerenzung M, Bachorz J, Lewandowski H, Schweitzer C, Schmitt C, Kirn A, Hénon P
Institut de Recherche en Hématologie et Transfuison, Hôpital du Hasenrain, Mulhouse, France.
Stem Cells. 1994 Mar;12(2):187-97. doi: 10.1002/stem.5530120207.
Circulating CD34+ progenitors were separated from normal human peripheral blood on the basis of size and density by counterflow centrifugal elutriation (CCE). The CD34+ cells, 0.15% of peripheral blood mononuclear cells, were heterogeneous with respect to their elutriation characteristics, mainly size and density. The least mature CD34+ cells, characterized by lack of CD38 antigen, were predominantly found in the small lymphoid cell fraction. In fractions containing larger and denser cells (large lymphocytes, monocytes, and granulocytes), CD38 was increasingly expressed on the CD34+ cells, as were lineage commitment markers CD10 (B lymphoid), CD33 (myeloid), CD13 (myelomonocytic) and CD71 (erythroid) antigens. The smaller and less dense CD34+ cells expressed CD34 antigen brightly while the larger and denser CD34+ cells expressed it dimly. The smaller and less dense CD34+ high cells failed to establish colony growth in short-term culture while the larger and denser CD34+low cells gave rise to high counts of colony forming units-granulocyte macrophage (CFU-GM). Physical separation on the basis of size and density by CCE differentiates between two main classes of steady-state CD34+ cells from normal human peripheral blood. The smaller and less dense CD34+high cells correspond to the earliest progenitors that express differentiation markers poorly but CD34 antigen brightly, do not give rise to short-term colony growth in vitro, and thus represent indirect evidence for pluripotent hematopoietic stem cells (PHSC). The larger and denser CD34+low cells are the more mature progenitor cells, already committed to myeloid, lymphoid or erythroid differentiation but only dimly expressing CD34 antigen, and these cells were responsible for short-term colony growth in vitro.
通过逆流离心淘析（CCE）根据大小和密度从正常人外周血中分离出循环CD34+祖细胞。CD34+细胞占外周血单个核细胞的0.15%，其淘析特征（主要是大小和密度）存在异质性。最不成熟的CD34+细胞缺乏CD38抗原，主要存在于小淋巴细胞部分。在含有较大且密度较高细胞（大淋巴细胞、单核细胞和粒细胞）的部分中，CD34+细胞上CD38的表达逐渐增加，谱系定向标记物CD10（B淋巴细胞）、CD33（髓系）、CD13（髓单核细胞）和CD71（红系）抗原的表达也增加。较小且密度较低的CD34+细胞CD34抗原表达明亮，而较大且密度较高的CD34+细胞则表达暗淡。较小且密度较低的CD34+高表达细胞在短期培养中未能形成集落生长，而较大且密度较高的CD34+低表达细胞产生了大量的集落形成单位-粒细胞巨噬细胞（CFU-GM）。通过CCE基于大小和密度进行物理分离可区分正常人外周血中稳态CD34+细胞的两个主要类别。较小且密度较低的CD34+高表达细胞对应于最早的祖细胞，其分化标记物表达较差但CD34抗原表达明亮，在体外不能产生短期集落生长，因此代表了多能造血干细胞（PHSC）的间接证据。较大且密度较高的CD34+低表达细胞是更成熟的祖细胞，已定向分化为髓系、淋巴系或红系，但仅微弱表达CD34抗原，这些细胞负责体外短期集落生长。
