造血生长因子对长期培养中骨髓造血细胞增殖和分化的影响

Effect of HGFs on proliferation and differatiation of HSCs in LTBMC

应用骨髓长期培养法研究了ＨＧＦｓ联合应用对ＬＴＢＭＣ中造血干／祖细胞的增殖、分化的影响，结果显示，在ＬＴＢＭＣ中，含有ＨＧＦｓ（ＩＬ３＋ＩＬ６＋ＧＣＳＦ＋ＧＭＣＳＦ＋Ｅｐｏ）的扩增组与对照组相比，ＨＧＦｓ能显著地扩增ＨＳＣｓ，在第一周，ＣＦＵＧＭ，ＣＦＵＥ和ＢＦＵＥ总数分别扩增了１８０９１６５９倍、１０２６８４８倍和１４９９１２７８倍，而累积扩增倍数分别达３６１９６６３６倍、２０５１３３９０倍和２９９８５１１３倍，有核细胞总数扩增了１６９１９８倍，累积扩增了３９６６９７倍，但２周后各种集落数及大型ＣＦＵＧＭ集落数下降明显加快。结果提示：ＨＧＦｓ组合应用结合长期培养能扩增造血干／祖细胞，并能提高悬浮细胞的生存率；ＨＧＦｓ的组合在扩增ＨＳＣｓ的同时也促进了其分化；对骨髓造血微环境的建立也具有促进作用，在适当的时间内，应用小容量骨髓单个核细胞在体外长期培养扩增体系中可持续地扩增，扩增的造血干／祖细胞具有重要的临床应用潜能。

The effects of long term bone marrow culture (LTBMC) containing the HGFs (IL 3+IL 6+G CSF+GM CSF+Epo) on proliferation and differentiation of hematopoietic progenitor cells in the culture have been studied. The findings showed that the LTBMC combining with human hematopoietic growth factors that include IL 3、IL 6、Epo、G CSF and GM CSF can expand the hematopoietic progenitor cells significantly. Compared with the untreated LTBMC containing no HGFs, the numbers of CFU GM、CFU E and BFU E were expanded 18 09 to 6 59、10 26 to 8 48 and 14 99 to 12 78 fold, respectively, the cumulative expansion of these colonies were 36 19 to 66 36、20 51 to 33 90 and 29 98 to 51 13 fold, respectively, during the first week. Similarly, the total number of nonadherent cells was expanded 1 69 to 1 98 fold and the cumulative expansion fold was 3 69 to 6 79 fold, but, after two weeks, the numbers of all kinds of colonies and the huge colonies formed from CFU GM were decrease rapidly, our data suggest that the combination of HGFs with LTBMC may promote the proliferation and differentiation of HSCs and establishment of bone marrow hematopoietic microenvironment. It is possible to expand small numbers of BM MNCS steadily in this LTBMC system within an appropriate peroid. Expansion of HSCs has a potential value of clinical application.