prikaz prve stranice dokumenta Kontrola kvalitete stanične terapije autolognim keratinocitima kod zacjeljivanja teških opeklina
  Download
PDF 2.28 MB
master's thesis
Kontrola kvalitete stanične terapije autolognim keratinocitima kod zacjeljivanja teških opeklina
Zagreb: University of Zagreb, Faculty of Science, 2021. urn:nbn:hr:217:349088
Arić, Ivona
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Arić, I. (2021). Kontrola kvalitete stanične terapije autolognim keratinocitima kod zacjeljivanja teških opeklina (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:349088

Arić, Ivona. "Kontrola kvalitete stanične terapije autolognim keratinocitima kod zacjeljivanja teških opeklina." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:349088

Arić, Ivona. "Kontrola kvalitete stanične terapije autolognim keratinocitima kod zacjeljivanja teških opeklina." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:349088

Arić, I. (2021). 'Kontrola kvalitete stanične terapije autolognim keratinocitima kod zacjeljivanja teških opeklina', Master's thesis, University of Zagreb, Faculty of Science, accessed 17 May 2024, https://urn.nsk.hr/urn:nbn:hr:217:349088

Arić I. Kontrola kvalitete stanične terapije autolognim keratinocitima kod zacjeljivanja teških opeklina [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2021 [cited 2024 May 17] Available at: https://urn.nsk.hr/urn:nbn:hr:217:349088

I. Arić, "Kontrola kvalitete stanične terapije autolognim keratinocitima kod zacjeljivanja teških opeklina", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2021. Available at: https://urn.nsk.hr/urn:nbn:hr:217:349088

Metadata
TitleKontrola kvalitete stanične terapije autolognim keratinocitima kod zacjeljivanja teških opeklina
Title (english)Quality control of cell therapy with autologous keratinocytes for severe burn wound healing
AuthorIvona Arić
MentorMarija Zekušić (mentor)
MentorInga Urlić (sumentor)
Committee memberInga Urlić (predsjednik povjerenstva)
Committee memberJasna Lajtner (član povjerenstva)
Committee memberSofia Ana Blažević (član povjerenstva)
Committee memberPetra Peharec Štefanić (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2021-02-25, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Lijekovi za naprednu terapiju (engl. Advanced Therapy Medicinal Products, ATMP) pomažu pri liječenju opeklina i rana koje teško zacjeljuju. U Banci tkiva i stanica u Kliničkom bolničkom centru Sestre milosrdnice uzgaja se i klinički primjenjuje autologna suspenzija keratinocita. Duboke dermalne opekline (II B°) i subdermalne opekline (III°) nemaju sposobnost spontanog zacjeljivanja zbog nedostataka keratinocitnih matičnih stanica. Nakon biopsije kože, enzimatskom digestijom se odvaja epidermis od dermisa, a zatim se izoliraju pojedinačni keratinociti i zasijavaju na 3T3 stanice tretirane mitomicinom C koje služe kao hranjiva podloga. Keratinociti se uzgajaju u plastičnim bočicama u mediju koji je obogaćen serumom i faktorima rasta. Klinički se primjenjuju u šprici sa dvokomponentnim fibrinskim ljepilom. Neophodna je kontrola kvalitete tijekom uzgoja koja podrazumijeva provjeru ukupnog broja keratinocita, postotak vijabilnih stanica i klonogeničnosti (holokon, meroklon, paraklon). Udio ostatnih 3T3 stanica u suspenziji određuje se metodom protočne citometrije. U staničnoj terapiji se imunocitokemijskim metodama provjerava prisutnost epitelnih stanica bazalnog porijekla (CK14) te matičnih (p63), proliferacijskih (Ki-67) i diferenciranih (CK19) stanica. Mikrobiološkim kontrolama se dokazuje sterilnost svih staničnih medija te se mjeri koncentracija bakterijskih endotoksina i provjerava odsutnost Mycoplasma spp. Kontrola kvalitete osigurava efikasnu i sigurnu primjenu autolognih keratinocita kao ATMP.
Abstract (english)
Advanced therapy medicinal products (ATMPs) help treat burns and wounds that are difficult to heal. An autologous keratinocyte suspension is cultivated at the Tissue and Cell Bank in Sestre milosrdnice University Hospital Center. Deep dermal burns (II B°) and subdermal burns (III°) do not have the ability to heal spontaneously due to keratinocyte stem cell deficiencies. After skin biopsy, enzymatic digestion separates the epidermis from the dermis and then individual keratinocytes are isolated and seeded onto mitomycin C treated 3T3 cells used as a nutrient medium. Keratinocytes are cultivated in flasks in a serum and growth factors enriched medium. Clinically, they are applied by a syringe using fibrin glue. A quality control during cultivation in necessary and it includes keratinocyte number count, viable cells percentange and clonogenicity (holoclon, meroclone, paraclone). A number of the rest of 3T3 cells in the suspension is determined using flow cytometry. In the cell therapy, immunocytochemic methods are used for determining the presence of basal epithelial cells (CK14) and stem (p63), proliferation (Ki-67) and differential (CK19) cells. Microbiological controls prove the sterility of all cell mediums as well as measure the concentration of bacterial endotoxins and check for Mycoplasma spp. absence. Quality control ensures efficient and safe autologous keratinocytes usage as ATMP.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:349088
Study programmeTitle: Molecular Biology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2023-02-25
Terms of useLicence In copyright icon
Created on2021-03-08 14:15:59