Primerjava lastnosti matičnih celic iz aspiratov foliklove tekočine, pridobljene v postopku oploditve z biomedicinsko pomočjo pri različno starih pacientkah

Omejec, Sandra (author), Virant Klun, Irma (mentor)

V aspiratih foliklove tekočine, pridobljene kot stranski produkt ob aspiraciji jajčnih celic iz foliklov, v postopku oploditve z biomedicinsko pomočjo, je prisotna podpopulacija celic, ki ima lastnosti matičnih celic. Predhodne študije so pokazale veliko heterogenost teh celic, kar je slabost za njihovo morebitno uporabo v terapevtske namene, saj morajo biti lastnosti takih celic natančno določene. Zato je bil naš namen raziskati vpliv starosti pacientk, vključenih v postopek oploditve z biomedicinsko pomočjo, na lastnosti matičnih celic iz aspiratov foliklove tekočine. Pacientke smo razdelili v štiri starostne skupine (A [< 31 let], B [31–35 let], C [36–40 let], D [40–43 let]) in primerjali lastnosti njihovih celic iz foliklove tekočine. Število celic iz aspiratov foliklove tekočine se statistično značilno ni razlikovalo glede na starost pacientk. Celice so bile vretenaste ali kubične oblike, morfologija celic pa ni bila povezana s starostjo pacientk. Prav tako se ni razlikoval delež osamljenih celic, ki so se pozitivno obarvale po imunocitokemijskembarvanju površinskih označevalcev, značilnih za mezenhimske matične celice (CD146, CD44 in CD90). Podrobnejša genetska analiza pa je pokazala zanimive razlike izražanja genov v celicah glede na starost pacientk.,Z analizo smo izpostavili gene, ki so bili različno izraženi ali so biliizraženi samo pri najmlajših pacientkah skupine A ali najstarejših pacientkah skupine D, vključno z geni, značilnimi za pluripotentnost; samo v celicah najmlajših pacientk sta bila izražena gena LIF in POU5F1, medtem ko je bil gen WNT3A, povezan s staranjem tkiv in nastankom raka, izražen samo v celicah najstarejših pacientk skupine D. Kljub temu, da smo ugotovili najvišje izražanje genov, značilnih za mezenhimske matične celice, so se celice najstarejših pacientk skupine D najslabše diferencirale v adipogene celice alise sploh niso diferencirale v osteogene celice. Rezultati tega dela dokazujejo vpliv starosti ženske na kakovost in funkcijo matičnih celic iz foliklove tekočine.

• Language: Slovenian
• Keywords: celice granuloze/ mezenhimske matične celice/ foliklova tekočina/ jajčnik/starost pacientk
• Work type: Master's thesis/paper
• Organization: BF - Biotechnical Faculty
• Year: 2017
• PID: 20.500.12556/RUL-97575
• COBISS.SI-ID: 4513359
• Publication date in RUL: 27.10.2017

Secondary language

• Language: English
• Title: Characterization of stem cell propreties of aspirated follicular cells derived from different aged patients

Abstract

Follicular aspirates obtained in clinical practice in assisted reproduction as a side product contain a subpopulation of cells with stem cell characteristic. Previous studies showed high degree of heterogeneity among such cells, which is a drawback for their potential use in therapeutic purposes where the cell characteristics need to be precisely defined. Therefore the aim of the study was to investigate the effect of age of patients included in the program of assisted reproduction on the stem cell characteristics of cells isolated from follicular aspirates. The patients were divided in four age groups (A [< 31 years], B [31–35 years], C [36–40 years], D [40 < years]) to compare the characteristics of isolated cells. The number and morphology of cells isolated from follicular aspirates was not statistically different among patients of different age groups.. The cells had the spindle shaped and cuboidal morphology in all age groups. Also the proportion of isolated cells positively stained for some mesenchymal stem cell surface markers (CD146, CD44 in CD90) using immunocytochemistry did not differ between age groups. Gene expression analysis showed interesting differential expression of genes that were expressed only in the youngest group age A or in the oldest group age D, including genes related to pluripotency; LIF and POU5F1 were expressed only in cells from the youngest patients, while WNT3A previously associated with tissue aging and cancer development was expressed only in the oldest group of patients. Although the highest expression of genes specific for mesenchymal stem cells was detected in the oldest group of patients D, their adipogenic differentiation was the least successful and they were not able to differentiate into osteogenic cells. The results of this study demonstrate the effect of female age on quality and function of stem cells from follicular fluid.

• Keywords: granulosa cells/ mesenchymal stem cells/ follicular fluid/ ovary/ age of patients