Zychowicz Marzena, Mehn Dora, Ruiz Ana, Colpo Pascal, Rossi Francois, Frontczak-Baniewicz Malgorzata, Domanska-Janik Krystyna, Buzanska Leonora
NeuroRepair Department, Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland.
Acta Neurobiol Exp (Wars). 2011;71(1):12-23. doi: 10.55782/ane-2011-1819.
Physical interactions of cells with the adhesive substrates of the microenvironment as well as the presence of the soluble growth factors are important for the proliferation capacity of neural stem cells. We have used biofunctionalized surface domains microcontact printed with either synthetic polyaminoacid poly-L-lysine or extracellular matrix (ECM) component such as fibronectin, to study the proliferation capacity of human umbilical cord blood-derived neural stem cells (HUCB-NSC). The proliferation measured by the expression of Ki-67 protein was accompanied by the investigation of the cell morphology under the transmission and scanning electron microscopy in different culture time, plating densities of cells and medium condition (serum-free or 2 percent of FBS). The poly-L-lysine domains of defined micro-scale area promoted the presence of round, loosely attached Ki-67-positive cells, while fibronectin domains of the same size allowed appearance of flattened, strongly attached cells with more differentiated phenotype. These results were in agreement with the non-specific, electrostatic type of interaction between cell and substrate on poly-L-lysine and integrin receptor-mediated specific adhesion on fibronectin. In this report we have described in vitro culture conditions, which allow for immobilization of the non-differentiated and highly proliferating population of neural stem/progenitor cells to the biofunctionalized surface. The microarrays with bioactive domains allocating non-differentiated and proliferating neural stem/progenitor cells may find application for drug and chemicals toxicology screening of diverse factors influencing neural development.
细胞与微环境的黏附底物之间的物理相互作用以及可溶性生长因子的存在对神经干细胞的增殖能力很重要。我们使用了用合成聚氨基酸聚-L-赖氨酸或细胞外基质（ECM）成分如纤连蛋白进行微接触印刷的生物功能化表面区域，来研究人脐带血来源的神经干细胞（HUCB-NSC）的增殖能力。通过Ki-67蛋白表达测量的增殖伴随着在不同培养时间、细胞接种密度和培养基条件（无血清或2%胎牛血清）下，利用透射和扫描电子显微镜对细胞形态的研究。特定微尺度区域的聚-L-赖氨酸区域促进了圆形、松散附着的Ki-67阳性细胞的存在，而相同大小的纤连蛋白区域则允许出现扁平、紧密附着且具有更分化表型的细胞。这些结果与细胞与聚-L-赖氨酸上的底物之间的非特异性静电相互作用以及纤连蛋白上整合素受体介导的特异性黏附一致。在本报告中，我们描述了体外培养条件，该条件允许将未分化且高度增殖的神经干/祖细胞群体固定在生物功能化表面上。带有分配未分化和增殖神经干/祖细胞的生物活性区域的微阵列可能会在影响神经发育的各种因素的药物和化学品毒理学筛选中找到应用。
