Eastment C E, Ruscetti F W, Denholm E, Katznelson I, Arnold E, Ts'o P O
Blood. 1982 Aug;60(2):495-502.
In contrast to the murine system, long-term hamster bone marrow suspension cultures maintain proliferation of both pluripotent and committed stem cells in the absence of an adherent layer and without addition of exogenous factors, such as hydrocortisone. Addition of pokeweed-mitogen-stimulated hamster spleen conditioned medium (SCM) to these long-term suspension cultures produces an increase in the number of mixed colonies assayed in soft-agar, These mixed colonies, which contained four cell lineages--granulocytic, erythroid, megakaryocytic, and macrophage--could be generated from cells grown in suspension for over 6 mo. Addition of SCM also induces an initial rapid expansion of the myeloid compartment, and this expansion results in 70% of the cells being terminally differentiated granulocytes. In contrast, addition of SCM to hamster bone marrow cultures containing both adherent cells and hematopoietic stem cells produced no change in the number of mixed colonies generated in the culture. This system allows the in vitro study of the process of stem cell proliferation and differentiation and also provides a means to examine the relationship of adherent and supernatant bone marrow populations.
与小鼠系统不同,长期仓鼠骨髓悬浮培养在没有贴壁层且不添加外源性因子(如氢化可的松)的情况下,能维持多能干细胞和定向干细胞的增殖。向这些长期悬浮培养物中添加商陆丝裂原刺激的仓鼠脾条件培养基(SCM),会使软琼脂中检测到的混合集落数量增加。这些混合集落包含粒细胞、红细胞、巨核细胞和巨噬细胞这四种细胞谱系,可从悬浮培养超过6个月的细胞中产生。添加SCM还会诱导髓系区室最初的快速扩增,这种扩增导致70%的细胞最终分化为粒细胞。相比之下,向含有贴壁细胞和造血干细胞的仓鼠骨髓培养物中添加SCM,培养物中产生的混合集落数量没有变化。该系统允许对干细胞增殖和分化过程进行体外研究,还提供了一种方法来研究贴壁和上清骨髓群体之间的关系。