WACHSTUMSTIMULIERENDES MATERIAL, HERSTELLUNGSVERFAHREN UND THERAPEUTISCHE ZUSAMMENSETZUNG.

摘要

Growth-stimulating materials (I) comprising mitogenic factor(s) are produced by (a) cooling defatted metacarpal bone and/or a bone mixt. from pigs in liq. N2; (b) comminuting the bone to a particle size of 0.5-2 mm in a press at the temp. of liq. N2; (c) demineralising the powder with an aq. soln. of a complexing agent; (d) dialysing, acidifying and heating the resulting aq. suspension; (e) centrifuging to remove insolubles; (f) opt. chromatographing the supernatant on hydroxyapatite, eluting with guanidine.HCl; and (g) opt. subjecting the eluate to gel filtration under high-salinity conditions to isolate a 24 +/- 8 kD fraction. (I) have the following properties: (a) produce dose-dependent growth stimulation of fibroblasts, osteoblast precursor cells and chondrocytes from chicken embryo tissue in monolayer cell cultures; (b) produce dose-dependent growth stimulation of chicken embryo vertebra in tissue culture, resulting in strong proliferation of chondrocytes in the growth zone; (c) do not increase 3H-thymidine uptake in Calvaria cell DNA; (d) have effects (a) and/or (b) lasting for at least 15 min. at 55-75 deg.C and for at least 20 min. at pH 3-10; (e) have a molecular wt. of 24 +/- 8 kD by gel filtration under high-salinity conditions; (f) pass unadsorbed through a CuSO4 chelate column, with a 5- to 20-fold increase in specific activity; and (g) can be bound on a reverse-phase HPLC column and eluted with 10-45% aq. acetone. Therapeutic compsns. comprising (I) and a solid carrier (II) are new.