Multiple emulsions for cryopreservation and banking of living cells
Agata Metera , Agnieszka Markowska-Radomska , Konrad Kosicki , Ewa Dłuska
AbstractThe paper presents the results of encapsulation and cryopreservation of living Human Embryonic Kidney cells (HEK) in multiple emulsions. Double emulsions have been proposed and discussed as prospective living cells carriers for cryopreservation and banking. Emulsions were prepared by one step method in a Couette-Taylor flow (CTF) biocontactor. W1/O/W2 emulsions consisted of alginic acid aqueous solution as inner phase, liquid paraffin as oil membrane phase, distilled water as outer phase. Appropriate surfactants were added to each phase of multiple emulsion. Sucrose as a cryoprotectant was added to inner phase of the emulsion and optionally to outer phase of freshly prepared emulsion before freezing. Cooling was carried out with freezing container to achieve a rate of cooling close to -1 degree Celsius per minute within a -80 degree Celsius refrigerator. Experimental study on the encapsulation and cryopreservation involved: the influence of process parameters in the CTF biocontactor on the living cells encapsulation efficiency and the influence of cryprotectant addition to outer phase of multiple emulsion on cells viability. W1/O/W2 emulsions with living cells encapsulated in the inner phase were obtained. Living cells encapsulation efficiency was close to 100% in the investigated double emulsion. Addition of sucrose to inner and outer phase of emulsion resulted in over 90% cells viability after thaw of the samples. Cell viability in samples with sucrose in both aqueous phases of emulsions remained almost constant for 47 days. Cells viability in samples with sucrose in only inner aqueous phase was over 55% for two days and then fallen to 23-20%. The obtained results regarding cells viability showed that multiple emulsions may be proposed as an innovative living cells carrier for cryopreservation and banking.
|Publication size in sheets||0.3|
|Book||Kędzierska K., Budniak U., Jancewicz I., Krupiński P., Kutkowska A., Monczarska D., Paziewska Agnieszka, Paziewska A., Spreavier A., Trzebuniak K. (eds.): V Międzyuczelnianie Sympozjum Biotechnologiczne „Symbioza” im. Prof. Krzysztofa W. Szewczyka Księga Abstraktów, 2016, Warszawa, Uniwersytet Warszawski, ISBN 9788394234232, , 192 p.|
|Keywords in English||multiple emulsions, helical contactor, cells encapsulation, cells banking|
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