Dresch C, Barreau P
Exp Hematol. 1985 Dec;13(11):1143-51.
In blood and marrow samples from ten normal adults, mixed colonies were found in agar cultures at concentrations of 4.1 +/- 3.4/10(5) mononuclear bone marrow cells, and of 4.7 +/- 4/5 X 10(5) mononuclear blood cells. Reseeding of five- to seven-day clusters of immature cells in fresh medium with 5% PHA leukocyte-conditioned medium (PHA-LCM) and 2 U erythropoietin (Epo) showed that 10(5) mononuclear cells contained as many as 42 +/- 12 CFU-mix in bone marrow and 12 +/- 7 CFU-mix in blood, i.e., ten times more than in the primary cultures. Attempts were made to discover why these potentially mixed colonies failed to develop in primary cultures. For this purpose five- to seven-day clusters of immature cells from cultures of 5 X 10(4) bone marrow cells grown with PHA-LCM, Epo, or both were reseeded in medium containing one or both stimulants. The presence of both in secondary culture gave the best CFU-mix growth, whatever the primary stimulation, i.e., more than one-third of the immature clusters gave rise to colonies of red cells and granulocyte lineages. The addition of one or both stimulants between days 7 and 14 of primary culture, however, did not increase the number of mixed colonies or give rise to late developing ones. Reseeding of colonies and clusters of 14-day primary cultures showed that at least one-third of erythroid bursts or of late-developing granulocyte colonies contained CFU-mix, but that less than 10% of 14-day clusters developed into mixed colonies. Cultures of nonadherent cells, mononuclear cells of less than 1.065/g density or with a velocity sedimentation rate of less than 6 mm/h gave the same results as the culture of total mononuclear cells; i.e., at least ten times more multipotent progenitors were found by reseeding after five to seven days than in primary cultures. We conclude that in human bone marrow and blood the physical and kinetic properties of pluripotent stem cells are similar to those of 14-day erythroid bursts and GM colonies. Under normal culture conditions, most of these cell types differentiate into one cell lineage only, and plurilineage differentiation is inhibited.
在来自10名正常成年人的血液和骨髓样本中，在琼脂培养物中发现混合集落，其浓度在每10⁵个骨髓单个核细胞中为4.1±3.4个，在每5×10⁵个血液单个核细胞中为4.7±4个。用5%PHA白细胞条件培养基（PHA-LCM）和2U促红细胞生成素（Epo）在新鲜培养基中对5至7天的未成熟细胞簇进行再接种，结果显示每10⁵个单个核细胞在骨髓中含有多达42±12个混合集落形成单位（CFU-mix），在血液中含有12±7个CFU-mix，即比原代培养中的数量多10倍。人们试图探究为什么这些潜在的混合集落在原代培养中未能发育。为此，将用PHA-LCM、Epo或两者培养的5×10⁴个骨髓细胞培养物中5至7天的未成熟细胞簇在含有一种或两种刺激物的培养基中进行再接种。无论原代刺激如何，在继代培养中同时存在这两种刺激物时CFU-mix生长最佳，即超过三分之一的未成熟细胞簇产生红细胞和粒细胞系的集落。然而，在原代培养的第7天至第14天添加一种或两种刺激物，并未增加混合集落的数量或产生晚期发育的集落。对14天原代培养的集落和细胞簇进行再接种显示，至少三分之一的红系爆式集落或晚期发育的粒细胞集落含有CFU-mix，但14天的细胞簇中不到10%发育成混合集落。对非贴壁细胞、密度小于1.065/g或沉降速度小于6mm/h的单个核细胞进行培养，其结果与对总单个核细胞进行培养的结果相同；即5至7天后再接种时发现的多能祖细胞数量至少比原代培养中的多10倍。我们得出结论，在人类骨髓和血液中，多能干细胞的物理和动力学特性与14天的红系爆式集落和粒-巨噬细胞集落相似。在正常培养条件下，这些细胞类型中的大多数仅分化为一个细胞系，多系分化受到抑制。
