Utjecaj lizata trombocita na biološke značajke mezenhimskih matičnih stanica uzgojenih in vitro

master's thesis

• Title (english): BIOLOGICAL PROPERTIES OF MESENCHYMAL STEM CELLS EXPANDED IN VITRO IN MEDIA WITH DIFFERENT HUMAN PLATELET LYSATE CONTENT
• Author: Ksenija Božinović
• Mentor: Mirna Golemović (mentor)
• Mentor: Inga Marijanović (mentor)
• Committee member: Inga Marijanović (predsjednik povjerenstva)
• Committee member: Mirna Golemović (član povjerenstva)
• Committee member: Nada Oršolić (član povjerenstva)
• Committee member: Vesna Benković (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2012-07-03, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

U ovom istraživanju uzgajane su mezenhimske matične stanice (MMS) izolirane iz koštane srži (KS) tri dobrovoljna davatelja. MMS ispoljavaju imunomodulatorna svojstva in vitro što im daje potencijalnu primjenu pri transplantaciji krvotvornih matičnih stanica i liječenju reakcije presatka protiv primatelja. Učestalost MMS u KS je vrlo mala te ih je za potrebe kliničke primjene neophodno umnožiti in vitro. Za umnažanje MMS korištene su kultivacijske podloge oplemenjene s različitim udjelima lizata ljudskih trombocita (LT): 10, 8, 6, 4 ili 2% LT. MMS su uzgajane tijekom pet ciklusa presađivanja. Na kraju svakog ciklusa je analizirana kinetika rasta stanica, morfologija te imunofenotip uzgajanih MMS in vitro. Nakon drugog ciklusa umnažanja ispitan je adipogeni potencijal MMS. Tijekom cijelog perioda uzgoja in vitro, u svim uvjetima uzgoja stanice su ispoljavale mezenhimske karakteristike; prihvaćale su se na plastičnu podlogu, poprimile vretenasti fibroblastni izgled, ispoljavale imunofenotip MMS i uspješno diferencirale u zrele adipocite. Utvrđeno je da različiti udjeli LT ne utječu na navedene biološke značajke MMS, ali da imaju značajan utjecaj na kinetiku rasta MMS. Stanična populacija uzgojena u uvjetu s većim udjelom LT u kultivacijskoj podlozi se više umnažala u odnosu na stanice uzgajane u prisustvu manjeg udjela LT. Vrijeme udvostručenja stanica bilo je obrnuto proporcionalno udjelu LT: što je bio veći udio, stanice su se brže udvostručile.

Abstract (english)

Mesenchymal stem cells (MSCs) show immunomodulatory properties in vitro that make them interesting in the field of cellular therapy. MSCs have been clinically applied to improve engraftment in hematopoietic stem cell transplantation and to treat acute graft versus host disease. Incidence of MSCs in bone marrow (BM) is very low and in vitro expansion of MSCs is necessary. In this study MSCs derived from BM of three healthy donors were expanded in vitro. Cultivation mediums enriched with different proportion of human platelet lysate (PL) were used to expand MSCs: 10, 8, 6, 4 or 2% PL. MSCs were expanded through five passages. Cell growth kinetics, morphology and immunophenotype of expanded MSCs were analyzed. Following second passage, a small proportion of cells were used for analysis of adipogenic potential of expanded cells. During the whole period of cultivation in vitro, MSCs continuously expressed mesenchymal characteristics in all culture coditions: they adhered to plastic surface showing fibroblast shape, they expressed mesenchymal immunophenotype and differentiated into adypocites. Different content of PL in culture medium had no effect on investigated biological characteristics of the MSCs, but did have significant impact on the growth kinetics of MSC. With higher content of PL, number of harvested cells increased. Cell doubling time was inversely proportional to the content of PL in the medium.

• Keywords: mezenhimske matične stanice; lizat trombocita; koštana srž
• Keywords (english): mesenchymal stem cells; platelet lysate; bone marrow
• Language: croatian
• URN:NBN: urn:nbn:hr:217:220296
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-04-28 12:16:17