Effects Of Physical Factors On Retina Muller Cell (RMC)

Effects of Physical Factors on Retina Muller Cell (RMC) Retinal Muller cells (RMC) are the major glia cells in retina, they have properties of both astrocytes and oligodendrocytes, and are very important in maintaining normal retinal function and structure such as to support neurons, form retinal -blood barrier, store and buffer K~+ and participate in neuron synapse transduction. In recent years, it has shown that RMC play an important role in pathological process of retina, such as mechanical and light injury, diabetic retinopathy, ischemia and hypoxia. All these impacts can evoke a series changes in RMC's morphology and function. RMC's role in retina pathology process has become a focus in recent years. In this study, we observe the effects of HMW, infrasound and other physical factors on the RMC in vitro primarily, in order to offer some new data for deep and systematical RMC study. Part I Establishment of RMC culture system It is a very efficient method to study RMC by in vitro culture system, with its relative simple cell types, controlled experimental conditions and rapid process. As the cellular components in retina are very complex and the culture process is time-consuming, it is not an easy job to get pure RMC in a short time. In our study, we stimulate the primary cultured RMC with spleen cell conditioned medium (SCCM) which were obtained by CoriA activated spleen cell. The result show that this SCCM can significantly stimulate RMC growth, there are many other cell types mixed with RMC primary, but they can be eliminated by passaged the primary RMC culture (3 times) and result a pure RMC population. By immunocytochemistry detected, more than 95% of the passaged RMC are GFAP positive. To improve the culture process and shorten experimental period, we purified RMC by a new method: one step digestion followed by filtered once and then, shaken by physical rotary. It is a more efficient purification method which can get pure RMC from primary culture. This study put a basis for later experiments. Part fl The effects of high power microwave on RMC High power microwave (HPW) is an important conceptual weapon and intensive study is going on abroad. It is a notable trend for us. The biological effects of HPW have become a pressure task in an future point. Retina is one of the microwave sensitive organs, previous epidemic investigation and experimental study have shown that microwave can induce neuron atrophy, degeneration and necrosis, microvessel convulsion, patched blooding and effusion at the retina hind polar, all these result in abnormal visual function. But the injured mechanism is obscure. In order to elucidate the mechanism by which HMW damage visual function, we observe the effects of high power, short time pulse microwave on the RMC. The microwave source was chosen according to parameters as: frequency / 9.3GHz, power I 500MW; pulse width I 2122nm and repeat frequency / 0.1Hz. After radiated for 30 times, the RMC viability was assayed by MTT to determine their proliferation, cellular death was detected by trypan blue and apoptosis figure were tested by in situ TUNEL detected. The results are: 1. Compared to the control group, the radiated group has declined proliferation ability after radiated 24h, 96h and 120h. 2. Both the radiated and control group has a similar viability and without any significant difference between them as shown by trypan blue. 3. The...