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Kost in pokostnica golenice ter sklepna ovojnica kolena kot potencialni vir mezenhimskih matičnih/stromalnih celic pri kadavrih
ID Haring, Gregor (Author), ID Zupan, Janja (Mentor) More about this mentor... This link opens in a new window

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Abstract
Tkiva kadavrskih darovalcev predstavljajo dragocen alternativni vir za izolacijo primarnih celic z lastnostmi, podobnimi mezenhimskim matičnim/stromalnim celicam (MSC). Vendar pa so lastnosti primarnih celic, ki izhajajo iz različnih tkiv in v različnih obdobjih po smrti, slabo prepoznane. V doktorski nalogi želimo identificirati optimalni vir tkiva izmed treh kolenskih in obkolenskih tkiv za izolacijo primarnih celic, z lastnostmi, podobnimi MSC, in opredeliti vpliv časa po smrti na lastnosti teh celic. Pri 32 darovalcih smo v različnih posmrtnih intervalih odvzeli tkiva iz subhondralne kosti, sinovijske ovojnice in pokostnice. Primarne celice smo ovrednotili z uporabo standardnih metod v pogojih in vitro, kot so sposobnost tvorbe kolonij, trilinijske diferenciacije, imunofenotipizacijo, ter z novimi analizami, kot je določitev izražanja genov za nedavno odkrite označevalce človeških skeletnih matičnih celic. Naši rezultati kažejo, da primarne celice, izolirane iz teh treh tkiv, ne kažejo razlik v lastnostih, ki so značilne za MSC. Odkrili pa smo večje izražanje gena za negativni označevalec skeletnih matičnih celic, CD146 v celicah kostnega mozga. Dokazali smo tudi, da je stopnja uspešnosti izolacije primarnih celic odvisna od posmrtnega intervala. Vendar pa smo ugotovili tudi, da celice sinovijske ovojnice in pokostnice, izolirane več kot 48 ur po smrti, kažejo boljši osteogeni in hondrogeni potencial kot pa celice istih tkiv, izolirane hitreje po smrti. Rezultati te doktorske naloge tako dokazujejo, da so kolenska in obkolenska tkiva kadavrskih darovalcev tudi 3 dni po smrti strateški vir primarnih celic z lastnostmi MSC za uporabo v regenerativni medicini.

Language:Slovenian
Keywords:mezenhimske matične/stromalne celice, kadavri, koleno, obkolenska tkiva, kost in kostni mozeg, sinovijska ovojnica, pokostnica, analiza v pogojih in vitro, označevalci človeških skeletnih matičnih celic.
Work type:Doctoral dissertation
Organization:MF - Faculty of Medicine
Year:2023
PID:20.500.12556/RUL-145275 This link opens in a new window
Publication date in RUL:15.04.2023
Views:517
Downloads:67
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Secondary language

Language:English
Title:Tibial bone, periosteum and knee synovium as potential source of mesenchymal stem/stromal cells in cadavers​
Abstract:
Donor tissues from cadavers represent a valuable alternative source for the isolation of primary cells with mesenchymal stem/stromal cells (MSC)-like properties. However, the properties of primary cells obtained from different tissues and at different time points after death are poorly understood. In this doctoral thesis, we aimed to identify the optimal tissue source among three knee and peri-knee tissues for the isolation of primary cells with MSC-like properties, and to determine the influence of time after death on the properties of these cells. Tissues from the subchondral bone, synovium and periosteum were harvested from 32 donors at various post mortem intervals. Primary cells were analysed using routine in vitro methods such as colony-forming unit fibroblast assay, trilinear differentiation, immunophenotyping, and novel analyses, such as gene expression of recently identified markers of human skeletal stem cells. Our results show that primary cells isolated from three tissues are similar in MSC-specific properties. However, we detected increased expression of the gene encoding negative marker of human skeletal stem cells, CD146, in bone marrow cells. We also showed that the success rate of primary cell isolation depends on the post mortem interval. However, we also found that cells from the synovium and periosteum isolated more than 48 hours after death had better osteogenic and chondrogenic potential compared to cells from the same tissues isolated faster after death. The results of this thesis demonstrate that knee and peri-knee tissues of cadaveric donors are a strategic source of primary cells with MSC-like properties that can be used in regenerative medicine as even three days after death.

Keywords:mesenchymal stem/stromal cells, cadaver, knee, peri-knee tissue, bone and bone marrow, synovium, periosteum, in vitro analysis, human skeletal stem cell markers.

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