Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN 55455, USA.
J Biotechnol. 2010 Oct 1;150(1):131-9. doi: 10.1016/j.jbiotec.2010.08.001. Epub 2010 Aug 7.
Many potential applications of stem cells require large quantities of cells, especially those involving large organs such as the liver. For such applications, a scalable reactor system is desirable to ensure a reliable supply of sufficient quantities of differentiation competent or differentiated cells. We employed a microcarrier culture system for the expansion of undifferentiated rat multipotent adult progenitor cells (rMAPC) as well as for directed differentiation of these cells to hepatocyte-like cells. During the 4-day expansion culture, cell concentration increased by 85-fold while expression level of pluripotency markers were maintained, as well as the MAPC differentiation potential. Directed differentiation into hepatocyte-like cells on the microcarriers themselves gave comparable results as observed with cells cultured in static cultures. The cells expressed several mature hepatocyte-lineage genes and asialoglycoprotein receptor-1 (ASGPR-1) surface protein, and secreted albumin and urea. Microcarrier culture thus offers the potential of large-scale expansion and differentiation of stem cells in a more controlled bioreactor environment.
许多干细胞的潜在应用需要大量的细胞，特别是那些涉及肝脏等大型器官的应用。对于此类应用，需要可扩展的反应器系统来确保可靠地供应足够数量的分化能力或分化细胞。我们使用微载体培养系统来扩增未分化的大鼠多能成体祖细胞（rMAPC），并将这些细胞定向分化为肝样细胞。在 4 天的扩增培养过程中，细胞浓度增加了 85 倍，而多能性标记物的表达水平以及 MAPC 的分化潜能保持不变。在微载体上定向分化为肝样细胞的效果与在静态培养中观察到的细胞相当。这些细胞表达了几种成熟的肝细胞谱系基因和去唾液酸糖蛋白受体-1（ASGPR-1）表面蛋白，并分泌白蛋白和尿素。因此，微载体培养为在更受控的生物反应器环境中大规模扩增和分化干细胞提供了潜力。
