Wang Bin, Kang Zi-zhen, Chi Zhan-you, Xu Li, Tan Wen-song
The State Key Laboratory of Bioreactor Engineering, ECUST, Shanghai 200237, China.
Zhonghua Xue Ye Xue Za Zhi. 2003 Feb;24(2):74-7.
To explore the ex vivo expansion characteristics of selected CD(34)(+) cells and mononuclear cells (MNC).
CD(34)(+) cells were isolated from umbilical cord blood MNC by MiniMACS system, expanded under the same conditions as that for MNC. The effects of re-isolation and the MNC supernatant (MNC-SN) on the selected CD(34)(+) cells were investigated. And the CD(34)(-) cells of MNC were cultured ex vivo.
In the culture of selected CD(34)(+) cells, both the colony density and the proportion of the CD(34)(+) cells declined continuously with the culturing, although they presented a high proliferation potential. However, in the culture of the MNC, from day 0 to day 7, the colony density and the proportion of CD(34)(+) cells were increased from 412 +/- 167/10(5) cells and (1.12 +/- 0.42)% to 1 162 +/- 566/10(5) cells and (4.17 +/- 1.44)%, respectively. It was found that both the total cells and the CD(34)(+) cells restored expansion potential by re-isolating. CD(34)(-) cells of MNC had the ability to form colony and could transform to CD(34)(+) cells. MNC-SN can promote colony forming ability and lead to CD(34)(+) cells differentiation at the same time.
In ex vivo culture, selected CD(34)(+) cells presented a high proliferation and differentiation potentials, and the CD(34)(-) cells produced during the cultivation had inhibition effect on CD(34)(+) cells expansion. CD(34)(-) cell population from cord blood MNC contained hematopoietic stem/progenitor cells and the cytokines secreted by CD(34)(-) cells could induce CD(34)(+) cells to more mature colony-forming cells.
探讨分选的CD(34)(+)细胞和单个核细胞(MNC)的体外扩增特性。
采用MiniMACS系统从脐带血单个核细胞中分离CD(34)(+)细胞,在与单个核细胞相同的条件下进行扩增。研究再次分选及单个核细胞上清液(MNC-SN)对分选的CD(34)(+)细胞的影响。并对单个核细胞的CD(34)(-)细胞进行体外培养。
在分选的CD(34)(+)细胞培养中,尽管其具有较高的增殖潜能,但随着培养的进行,集落密度和CD(34)(+)细胞比例均持续下降。然而,在单个核细胞培养中,从第0天到第7天,集落密度和CD(34)(+)细胞比例分别从412±167/10(5)个细胞和(1.12±0.42)%增加到1 162±566/10(5)个细胞和(4.17±1.44)%。发现通过再次分选,总细胞和CD(34)(+)细胞均恢复了扩增潜能。单个核细胞的CD(34)(-)细胞具有形成集落的能力,并可转化为CD(34)(+)细胞。MNC-SN可促进集落形成能力,同时导致CD(34)(+)细胞分化。
在体外培养中,分选的CD(34)(+)细胞具有较高的增殖和分化潜能,培养过程中产生的CD(34)(-)细胞对CD(34)(+)细胞的扩增有抑制作用。脐带血单个核细胞中的CD(34)(-)细胞群体含有造血干/祖细胞,CD(34)(-)细胞分泌的细胞因子可诱导CD(34)(+)细胞向更成熟的集落形成细胞分化。