Nahmias Yaakov, Kramvis Yiannos, Barbe Laurent, Casali Monica, Berthiaume Francois, Yarmush Martin L
Center for Engineering in Medicine/Department of Surgery, Massachusetts General Hospital, Shriners Burns Hospital, Harvard Medical School, Boston, MA 02114, USA.
FASEB J. 2006 Dec;20(14):2531-3. doi: 10.1096/fj.06-6192fje. Epub 2006 Oct 31.
Oxygen is an important component of the cellular microenvironment, mediating cell survival, differentiation, and function. Oxygen supply is a limiting factor during culture of highly metabolic cells such as hepatocytes. Here we present a simple formulation of a fluorocarbon-based oxygen carrier embedded in collagen gel that increases oxygen concentration in culture 6-fold. Rat hepatocytes cultured on oxygen carrier-collagen showed a significant increase in viability and function. Cytochrome P450IA1 activity was increased by 140% in serum-free cultures and by 820% in serum-containing cultures. The significantly higher hepatocellular function on oxygen carrier-collagen matrix persisted and increased during long-term culture. Long-term albumin secretion was increased by 350% in serum-free cultures and by 166% in serum-containing culture. Long-term urea secretion was increased by 79% in serum-free cultures and by 76% in serum-containing cultures. We conclude that oxygen supply may limit hepatocyte function in vitro. This limitation can be overcome by addition of an oxygen carrier to the extracellular matrix. Culture of hepatocytes on oxygen-carrying matrix mimics the oxygen-rich environment of the liver and provides a simple method for enhanced long-term function.
氧气是细胞微环境的重要组成部分，介导细胞存活、分化和功能。对于肝细胞等高代谢细胞的培养，氧气供应是一个限制因素。在此，我们展示了一种嵌入胶原凝胶中的基于氟碳化合物的氧载体的简单配方，其可使培养中的氧气浓度提高6倍。在氧载体 - 胶原上培养的大鼠肝细胞在活力和功能方面显著增加。在无血清培养中，细胞色素P450IA1活性提高了140%，在含血清培养中提高了820%。在氧载体 - 胶原基质上肝细胞功能显著更高，且在长期培养过程中持续存在并增加。在无血清培养中，长期白蛋白分泌增加了350%，在含血清培养中增加了166%。在无血清培养中，长期尿素分泌增加了79%，在含血清培养中增加了76%。我们得出结论，氧气供应可能会限制体外肝细胞功能。通过向细胞外基质中添加氧载体可以克服这种限制。在载氧基质上培养肝细胞模拟了肝脏的富氧环境，并提供了一种增强长期功能的简单方法。
