Fraunhofer Institute for Cell Therapy and Immunology, Perlickstraße 1, 04103, Leipzig, Germany.
J Inflamm (Lond). 2012 Apr 2;9(1):12. doi: 10.1186/1476-9255-9-12.
Studying primary adult microglia is hampered because of the difficult isolation procedure and the low cell yield. We therefore established a differentiation protocol using a culture system developed for the expansion of non-adherent bone marrow cells.
Non-adherent bone marrow derived stem cells (NA-BMC) are derived by selective adhesion ('preplating') and are non adhesive adult stem cells. We investigated the changes in bone marrow cell populations by this repeated selective adhesion and compared the potential of the derived cells to differentiate towards microglia. Cells were differentiated with astrocyte conditioned medium (ACM) and granulocyte-monocyte colony stimulating factor (GM-CSF).
NA-BMC cultures show a steep raise in the fraction of stem cells during the cultivation time and the differentiation potential is of the same quality as established protocols. Around 70% of the cells are microglia defined as being positive for CD11b/CD45 and show phagocytosis activity and oxidative bursts.
The non-adherent cell system has the advantage that is produces stem cell progenitors during expansion and provides good microglial differentiation.
由于原发性成年小胶质细胞的分离过程复杂且细胞产量低，因此对其进行研究受到了阻碍。我们因此建立了一种使用非贴壁骨髓细胞培养系统来扩增细胞的分化方案。
非贴壁骨髓来源的干细胞（NA-BMC）通过选择性黏附（“预培养”）得到，它们是非黏附的成体干细胞。我们通过这种反复的选择性黏附来研究骨髓细胞群的变化，并比较衍生细胞向小胶质细胞分化的潜力。细胞用星形胶质细胞条件培养基（ACM）和粒细胞-巨噬细胞集落刺激因子（GM-CSF）进行分化。
NA-BMC 培养物在培养过程中显示出干细胞比例的急剧上升，并且分化潜能与已建立的方案具有相同的质量。大约 70%的细胞为小胶质细胞，其特征为 CD11b/CD45 阳性，并表现出吞噬作用和氧化爆发。
非贴壁细胞系统的优势在于在扩增过程中产生干细胞祖细胞，并提供良好的小胶质细胞分化。
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