Amniotic membrane as a scaffold for human adipose stem cells and umbilical vein endothelial cells
Zhou, Mia (2018)
2018
Lääketieteen lisensiaatin tutkinto-ohjelma - Licentiate's Degree Programme in Medicine
Lääketieteen ja biotieteiden tiedekunta - Faculty of Medicine and Life Sciences
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Hyväksymispäivämäärä
2018-01-17
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi:uta-201802091199
https://urn.fi/URN:NBN:fi:uta-201802091199
Tiivistelmä
The amniotic membrane (AM) has many desirable qualities for tissue engineering. The three main objectives of this study were to investigate 1) adipose-derived stem cell (ASC) cultivation and possible differentiation on AM, 2) co-cultivation of ASCs and human umbilical vein endothelial cells (HUVECs) on AM for enhanced angiogenesis, 3) effects of cell cultivation on the mechanical properties of AM.
Denuded AM was cut and placed to cover the bottom of 6-well multidishes, 1/3 of AM-covered wells were seeded with only ASCs, 1/3 with only HUVECs and 1/3 with both. Half were cultured with adipose medium (ADM) and half with endothelial medium for three weeks (EM).
After three weeks, the cultivated AMs were analyzed with quantitative real-time polymerase chain reaction (RT-PCR), immunofluorescence photography, hematoxylin-eosin staining and microtomography. Tensile strength tests were conducted with separate AM samples cultivated in special scaffolds along with nitrocellulose paper.
Results showed no visible adipose formation, nor clear angiogenic tube formation. Engineering stress endured by AM during tension testing decreased as cultivation time lengthened. ASCs showed great viability on AM. HUVECs showed viability in EM based on von Willebrand factor expression. HUVEC proliferation was not seen in ADM. In conclusion, AM is a suitable scaffold for ASCs, better induction agents for adipocyte differentiation and tube formation should be explored, and a better method to detect HUVECs from the co-cultivation wells is needed.
Denuded AM was cut and placed to cover the bottom of 6-well multidishes, 1/3 of AM-covered wells were seeded with only ASCs, 1/3 with only HUVECs and 1/3 with both. Half were cultured with adipose medium (ADM) and half with endothelial medium for three weeks (EM).
After three weeks, the cultivated AMs were analyzed with quantitative real-time polymerase chain reaction (RT-PCR), immunofluorescence photography, hematoxylin-eosin staining and microtomography. Tensile strength tests were conducted with separate AM samples cultivated in special scaffolds along with nitrocellulose paper.
Results showed no visible adipose formation, nor clear angiogenic tube formation. Engineering stress endured by AM during tension testing decreased as cultivation time lengthened. ASCs showed great viability on AM. HUVECs showed viability in EM based on von Willebrand factor expression. HUVEC proliferation was not seen in ADM. In conclusion, AM is a suitable scaffold for ASCs, better induction agents for adipocyte differentiation and tube formation should be explored, and a better method to detect HUVECs from the co-cultivation wells is needed.