Optimal transportation protocol for human retinal pigment epithelium cell transportation on biomaterial membrane
Pietiläinen, Anna-Kaisa (2014)
2014
Materiaalitekniikan koulutusohjelma
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Hyväksymispäivämäärä
2014-06-04
Julkaisun pysyvä osoite on
http://urn.fi/URN:NBN:fi:tty-201405281248
http://urn.fi/URN:NBN:fi:tty-201405281248
Tiivistelmä
Ophthalmology group of the Institute of Biosciences and Medical Technology (Bio-MediTech) is a research group concentrating on studying retinal pigment epithelium cell (RPE) and their possibilities in cure of degenerative eye diseases. In the near future Ophthalmology group will do animal tests in co-operation with an European partner. For that reason RPE cells will be sent from Tampere to Germany.
Tissue banks have a wide experience of tissue storage and delivery. Regea is a cell and tissue center, which co-operates intensively with BioMediTech and is one of the Finnish tissue banks. Cells and tissues are sent weekly from Regea inside borders of Finland. Transportations take only a few hours. Many cell types and tissues are trans-ported either frozen or in hypothermal conditions. According to literature hypothermal conditions might not be the best choice for retinal pigment epithelium cells especially if transportation is over night.
Target of this thesis work was to formulate an optimal transportation protocol for human RPE cells on membrane material. Demonstrated transportations and cell ana-lysing methods were tested. Experimental part of this thesis was done with real RPE cells. Because cells and membranes are expensive, experiments were done with a small amount of samples. By using reasonable experiments it was possible to find optimal transportation conditions without use of statistical methods.
Results show that after 24 hours transportation RPE cells transported in room temperature in Hibernate A medium survived best. Immunohistochemical methods gave information about RPE cells and were suitable for cell analysis. Measurements of pH and transepithelial resistance are not recommended because of their low level of infor-mation in this experiment. The role of mechanical stress was higher than expected. Laboratory environment did not mimic air transportation exactly so there might be some unexpected results in the real transportation.
Tissue banks have a wide experience of tissue storage and delivery. Regea is a cell and tissue center, which co-operates intensively with BioMediTech and is one of the Finnish tissue banks. Cells and tissues are sent weekly from Regea inside borders of Finland. Transportations take only a few hours. Many cell types and tissues are trans-ported either frozen or in hypothermal conditions. According to literature hypothermal conditions might not be the best choice for retinal pigment epithelium cells especially if transportation is over night.
Target of this thesis work was to formulate an optimal transportation protocol for human RPE cells on membrane material. Demonstrated transportations and cell ana-lysing methods were tested. Experimental part of this thesis was done with real RPE cells. Because cells and membranes are expensive, experiments were done with a small amount of samples. By using reasonable experiments it was possible to find optimal transportation conditions without use of statistical methods.
Results show that after 24 hours transportation RPE cells transported in room temperature in Hibernate A medium survived best. Immunohistochemical methods gave information about RPE cells and were suitable for cell analysis. Measurements of pH and transepithelial resistance are not recommended because of their low level of infor-mation in this experiment. The role of mechanical stress was higher than expected. Laboratory environment did not mimic air transportation exactly so there might be some unexpected results in the real transportation.