Staber F G, Burgess A W
J Cell Physiol. 1980 Jan;102(1):1-10. doi: 10.1002/jcp.1041020102.
Serum from mice treated with bacterial lipopolysaccharide (LPS) was fractionated by Con A-Sepharose affinity chromatography, and assayed in vitro for colony-stimulating factor (CSF) using mouse bone marrow cells. The CSF failing to bind to concanavalin A-Sepharose (pool A) had similar biological properties to the unfractionated serum, i.e., it stimulated the formation of about equal numbers of granulocytic, mixed granulocyte-macrophage and macrophage colonies. The fraction eluted from the Con A-Sepharose column with alpha-methyl-D-glucopyranoside (pool B) had a steeper dose-response curve than either the unfractionated serum or the pool A CSF and most of the colonies were composed of macrophages. A mixture of the pool A and pool B CSFs stimulated colonies in a similar way as unfractionated serum and poolA. The apparent molecular weights of the two types of CSF were determined by two different gel-filtration procedures. Sephacryl S-200 gel-filtration suggested an apparent molecular weight of 85,000 for pool A CSF and 180,000 for pool B CSF. Gel-filtration on Sepharose CL-6B in the presence of guanidine hydrochloride (6M) yielded an apparent molecular weight of approximately 23,000 for pool A CSF and 33,000 for pool B CSF. The colony-forming cells (CFC) responding to pool B CSF were found to have a relatively high sedimentation velocity (peak sedimentation velocity 5.6--6.2 mm/hr) compared to the CFC responding to mouse-lung conditioned medium (MLCM) whose peak sedimentation velocity was between 4.0--4.5 mm/hour. The CFC responding to pool A CSF had an intermediate sedimentation velocity (peak 4.6--5.2 mm/hour). A time-course analysis of the morphology of clones or colonies in cultures stimulated with either MLCM or pool B CSF showed that the proportion of different colony types depends significantly on the incubation period and suggested that pool tb csf induced an early commitment of CFC towards macrophages differentiation.
用细菌脂多糖（LPS）处理过的小鼠血清经刀豆球蛋白A-琼脂糖亲和层析进行分级分离，并用小鼠骨髓细胞在体外检测集落刺激因子（CSF）。未与刀豆球蛋白A-琼脂糖结合的CSF（组分A）具有与未分级血清相似的生物学特性，即它刺激形成数量大致相等的粒细胞集落、粒细胞-巨噬细胞混合集落和巨噬细胞集落。用α-甲基-D-吡喃葡萄糖苷从刀豆球蛋白A-琼脂糖柱上洗脱下来的组分（组分B）比未分级血清或组分A的CSF具有更陡的剂量反应曲线，并且大多数集落由巨噬细胞组成。组分A和组分B的CSF混合物刺激集落的方式与未分级血清和组分A相似。通过两种不同的凝胶过滤方法测定了两种类型CSF的表观分子量。Sephacryl S-200凝胶过滤显示组分A的CSF表观分子量为85,000，组分B的CSF表观分子量为180,000。在6M盐酸胍存在下于Sepharose CL-6B上进行凝胶过滤，组分A的CSF表观分子量约为23,000，组分B的CSF表观分子量为33,000。与对小鼠肺条件培养基（MLCM）有反应的集落形成细胞（CFC）相比，对组分B的CSF有反应的CFC具有相对较高的沉降速度（峰值沉降速度5.6 - 6.2毫米/小时），对MLCM有反应的CFC的峰值沉降速度在4.0 - 4.5毫米/小时之间。对组分A的CSF有反应的CFC具有中等沉降速度（峰值4.6 - 5.2毫米/小时）。对用MLCM或组分B的CSF刺激的培养物中的克隆或集落形态进行的时间进程分析表明，不同集落类型的比例显著取决于孵育时间，并表明组分B的CSF诱导CFC早期向巨噬细胞分化的定向。
