Hinze Arnd, Stolzing Alexandra
Fraunhofer Institute for Cell Therapy and Immunology (IZI), Perlickstrasse 1, 04103, Leipzig, Germany.
BMC Cell Biol. 2011 Aug 19;12:35. doi: 10.1186/1471-2121-12-35.
Microglia, the macrophages of the brain, have been implicated in the causes of neurodegenerative diseases and display a loss of function during aging. Throughout life, microglia are replenished by limited proliferation of resident microglial cells. Replenishment by bone marrow-derived progenitor cells is still under debate. In this context, we investigated the differentiation of mouse microglia from bone marrow (BM) stem cells. Furthermore, we looked at the effects of FMS-like tyrosine kinase 3 ligand (Flt3L), astrocyte-conditioned medium (ACM) and GM-CSF on the differentiation to microglia-like cells.
We assessed in vitro-derived microglia differentiation by marker expression (CD11b/CD45, F4/80), but also for the first time for functional performance (phagocytosis, oxidative burst) and in situ migration into living brain tissue. Integration, survival and migration were assessed in organotypic brain slices.
The cells differentiated from mouse BM show function, markers and morphology of primary microglia and migrate into living brain tissue. Flt3L displays a negative effect on differentiation while GM-CSF enhances differentiation.
We conclude that in vitro-derived microglia are the phenotypic and functional equivalents to primary microglia and could be used in cell therapy.
小胶质细胞作为大脑中的巨噬细胞，与神经退行性疾病的病因有关，并且在衰老过程中表现出功能丧失。在整个生命过程中，小胶质细胞通过驻留小胶质细胞的有限增殖得以补充。骨髓来源的祖细胞对小胶质细胞的补充作用仍存在争议。在此背景下，我们研究了小鼠小胶质细胞从骨髓（BM）干细胞的分化情况。此外，我们还观察了FMS样酪氨酸激酶3配体（Flt3L）、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）对向小胶质样细胞分化的影响。
我们通过标志物表达（CD11b/CD45、F4/80）评估体外诱导的小胶质细胞分化情况，同时首次评估其功能表现（吞噬作用、氧化爆发）以及在原位向活脑组织的迁移能力。在脑片培养模型中评估细胞的整合、存活和迁移情况。
从小鼠骨髓分化而来的细胞表现出原代小胶质细胞的功能、标志物和形态，并能迁移到活脑组织中。Flt3L对分化有负面影响，而GM-CSF则增强分化。
我们得出结论，体外诱导的小胶质细胞在表型和功能上等同于原代小胶质细胞，可用于细胞治疗。
