Fractionation of human bone marrow cell suspensions in nylon fiber columns: an efficient method for the removal of cells that produce colony stimulating factor (CSF).
In this report, we describe an efficient technique for the extraction of CSF-producing cells from human marrow suspensions. Prior to plating in agar cultures, we incubated buoyant human marrow cells for 45 min in columns packed with nylon fiber or subjected the cells to two one-hour incubations in glass petri dishes. Recoveries of total cells, differential marrow elements, and committed granulocyte-monocyte progenitor cells (CFUc) were similar after each separative procedure. However, spontaneous CFUc proliferation was more effectively eliminated when cells were fractionated in nylon fiber columns. After the removal of cells which were adherent to glass, spontaneous CFUc proliferation in cultures containing no exogenous CSF accounted for 2.1% of total CFUc at a plating concentration of 10(5) cells/ml and 7.8% at a concentration of 3 X 10(5) cells/ml. After the fractionation of marrow cell suspensions in nylon fiber columns, spontaneous CFUc growth was completely obliterated at a plating concentration of 10(5) cells/ml, and at a concentration of 3 X 10(5) cells/ml accounted for only 0.09% of total CFUc. Further experiments were undertaken which demonstrated that buoyant marrow cells after incubation in nylon fiber columns may be employed to assay CSF in extremely dilute concentrations. Because of the simplicity and efficiency of this procedure, nylon fiber chromatography appears to be a highly useful technique for the rapid semi-purification of marrow suspensions for use in the assay of human CSF.
D001853:Bone Marrow / Q000378:metabolism; D001854:Bone Marrow Cells; D002469:Cell Separation / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods*; D003115:Colony-Stimulating Factors / Q000378:metabolism*; D006801:Humans; D009757:Nylons
D003115:Colony-Stimulating Factors; D009757:Nylons