Electrophysiological Study of Freshly Isolated Articular Chondrocytes Vs. Cryopreserved Chondrocytes

Upasana, Kachroo (2015) Electrophysiological Study of Freshly Isolated Articular Chondrocytes Vs. Cryopreserved Chondrocytes. Masters thesis, Christian Medical College, Vellore.

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Abstract

Cryopreservation of articular chondrocytes has recently gained ground because of its applications in cell culture, tissue engineering and reconstructive surgery. Questions may be raised as to the use of cryopreserved chondrocytes since this intervention may cause changes in chondrocyte phenotype and biology. One way of assessing phenotype maintenance is by studying the electrophysiological profile using the patch clamp technique. This study sought to compare freshly isolated and cryopreserved chondrocytes by recording ionic currents using patch clamp technique. Goat articular chondrocytes were isolated from cartilage shavings by enzyme digestion. Cell aliquots were transferred to liquid nitrogen after overnight gradual cooling to -80° C. Cells were cryopreserved for a period of 7-15 days (2 study groups- Day 7-10 and Day 11-15) Percentage viability was checked upon thawing and cells subjected to patch clamp analysis. Ionic currents were recorded in whole cell configuration using depolarizing potentials (VHold:-80mV; Test pulses: -80mV to +70mV at 10mV increments). Outwardly rectifying currents were recorded in fresh chondrocytes (n=6) and in cryopreserved cells (n=9+6) Comparison of current densities at all potentials above the threshold, revealed no significant difference between fresh and cryopreserved chondrocytes (p>0.05) Currents were blocked by 10mM TEA+ (a specific potassium channel blocker). Reversal potential for these currents was observed to be near the calculated equilibrium potential for potassium. The results indicate that chondrocytes remain viable and maintain ion channel functionality even after 15 days of cryopreservation. Cryopreserved chondrocytes may be utilized as an alternative when the use of freshly isolated chondrocytes poses a limitation.

Item Type: Thesis (Masters)
Uncontrolled Keywords: Freshly Isolated Articular Chondrocytes ; Cryopreserved Chondrocytes ; Electrophysiological study.
Subjects: MEDICAL > Physiology
Depositing User: Punitha K
Date Deposited: 24 May 2018 17:49
Last Modified: 27 May 2018 11:47
URI: http://repository-tnmgrmu.ac.in/id/eprint/8077

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