狗骨髓长期培养的贴壁层细胞及条件培养液对粒系祖细胞的影响

本文报告狗骨髓长期培养的初步研究结果。采用马血清与狗血清混合加入培养体系中,可建立起比较稳定的贴壁细胞层,可以维持19周以上。但是,这种贴壁层不能有效地维持造血干细胞的增殖与分化,上清液中血细胞与GM-CFU_C的数目,在第二次接种骨髓后2—3周即迅速下降。狗骨髓培养的条件液中存在着抑制GM-CFU_C生长的物质,但它对小鼠骨髓GM-CFU_C和CFU-S无影响。如以贴壁细胞作为底层,用双层琼脂平皿培养法与正常狗骨髓共同培养,发现贴壁细胞不仅没有抑制反而有加强CSF刺激GM-CFU_C生长的作用。因此对狗骨髓长期培养体系不能长期支持造血干细胞生长和繁殖的可能原因尚须作进一步探索。

EFFECT OF ADHERENT CELLS AND CONDITIONED MEDIUM OF LONG-TERM CANINE BONE MARROW CULTURE ON THE GRANULOCYTE/MACROPHAGE PROGENITOR CELLS

On the basis of murine and humanliquid long-term bone marrow cultures,we have successfully established theadherent cell layer in canine bonemarrow culture by addition of horseand dog serum mixture to the culturesystem.Although the adherent layerthus formed could be maintained aslong as 19 weeks,it could not effectivelysupport proliferation and differentia-tion of the haemopoietic stem cells inthe culture.Two to three weeks afterrecharging the fresh bone marrow cellson the adherent cell layer,the numberof nucleated cells and GM-CFUc inthe supernatant declined rapidly.Pre-liminarily experimental analyses demon-strated that some substance inhibitoryto the growth of GM-CFUc was presentin the conditioned medium of the caninebone marrow culture,but it did notaffect the GM-CFUc and CFU-S ofmurine bone marrow origin.By meansof double layer agar culture methodand using the adherent cells as feederlayer,the growth of colonies in theupper layer which contains fresh bonemarrow cells from the dog and CSFappears to be enhanced in comparisonwith the control in which the feederlayer contains no adherent cells.It isapparent that adherent cells do notproduce any inhibitory substance ag-ainst the growth of colonies,but on thecontrary,some substance(s) of thenature of potentiating the effect of CSFon the growth of colonies may dislplayitself.Although the presence of bothinhibitory and stimulating substancesin the culture system of dog bone mar-row,the cause of interference withhaemopoietic activity in vitro remainsto be investigated.