2016年3月28日星期一

Optical Oxygen Sensors for Applications in Microfluidic Cell Culture

Oxygen is an immensely important species in biological systems. Molecular oxygen plays a crucial role in the behavior and viability of many types of cells as well as the properties of human tissues.

Although the atmospheric oxygen level in air is 21%, the normal level in the human alveoli is 14%.

This level decreases away from the blood vessels and forms an oxygen gradient in many tissues, with normal levels varying from organ to organ. Hypoxia, or inadequate oxygen levels, has a large effect on cells and tissues, including inducing vasodilation and changing metabolic processes to reduce oxygen consumption. Tissue hypoxia in cancerous tumors has been linked with resistance to radiation therapy and many anticancer drugs, as well as increased likelihood of metastasis and decreased likelihood of patient survival and treatability. Oxygen levels in tumors are often significantly lower than those in normal tissues, leading to the development of hypoxia-activated anticancer drugs designed to specifically target the hypoxic tumor tissues.

Oxygen level has also been identified as an important parameter in stem cell cultivation and differentiation. Stem cell proliferation can be enhanced and apoptosis reduced in cultivation conditions with oxygen levels lower than the standard 20%. Changes in stem cell cultivation environment oxygen concentration can also be used to simulate in vitro the effects of disease.

Stem cell differentiation patterns are also highly dependent on oxygen levels. Embryonic development often occurs in low-oxygen environments, and oxygen has been found to be an important signal molecule to regulate stem cell differentiation. As such, carefully controlling the oxygen concentrations in stem cell populations in vitro is essential for controlling the cells’ differentiation and maintaining undifferentiated populations. In regenerative medicine, the transplantation of new stem cells may

be used to replace cells which have been lost through disease or injury. Understanding the dynamic oxygen conditions during normal tissue development will be necessary to control differentiation or apoptosis of stem cells. Oligodendrocyte progenitor cells, which may be used for the treatment of demyelinating diseases, should be initially cultured in 5% O2 and then differentiated in 20% O2 for increased cell production. These conditions should be reproduced in the production of cells for replacement therapies.

Because of the profound effect oxygen has on biological systems, controlling and monitoring oxygen concentrations is useful in many cell culture applications. Consequently, there has been much interest in the development of inexpensive oxygen sensors and control mechanisms that can be easily integrated with cell culture environments. In addition to the simple oxygen-sensing application, oxygen sensors can also be adapted for the measurement of glucose concentrations through the addition of glucose oxidase, which allows glucose levels to be determined from oxygen levels because an amount of oxygen dependent on the glucose concentration is consumed in the oxidation of glucose by glucose oxidase; this further increases the applicability of oxygen sensors.

Much of the early work on oxygen sensors focused on Clark-type electrode sensors, which detect a current flow caused by reduction of oxygen. Such sensors have been miniaturized and integrated with microfluidic devices to monitor the oxygen consumption of bacteria. The miniaturization of such devices requires microscale electrodes, and this type of sensor consumes oxygen (and thus requires sample stirring for accurate measurements), is easily contaminated by sample contents, and requires electrical connection between the sensor electrodes and the measurement infrastructure. These factors present several significant disadvantages for microfluidic cell culture systems.

Consequently, there has been much interest in the integration of optical oxygen sensors with microfluidic systems. These optical sensors present the advantages that they are easily miniaturized, are not easily contaminated, do not require physical contact between the sensor and optical detector, and do not consume oxygen. Most optical oxygen sensors operate on the principle of reversible luminescence quenching of the intensity or excited-state lifetime (as cited in) of a luminescent indicator dye or luminophore. This process occurs when the excited state energy of a fluorescent or phosphorescent indicator molecule is transferred to another molecule such as oxygen rather than being emitted in the form of a luminescence photon.




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