Developing an in vitro Stroke Model for Studying Adipose Stem Cells' Paracrine Effect on Neuronal Recovery After Ischemia
Nousiainen, Sonja (2019)
2019
Lääketieteen ja terveysteknologian tiedekunta - Faculty of Medicine and Health Technology
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Hyväksymispäivämäärä
2019-06-24
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi:tuni-201907092525
https://urn.fi/URN:NBN:fi:tuni-201907092525
Tiivistelmä
ABSTRACT
Stroke is one of the leading causes of mortality worldwide. Studies aiming to find new therapeutic methods improving patient’s outcome afterwards are conducted in the field of stem cell research. Adipose stem cells (ASC) may possess neuroprotective ability and they have shown a potential for helping neuronal cells suffered from ischemic injury to regenerate. The aim of the present research is to develop an in vitro ischemia model for studying ASC possible paracrine effect on damaged neuronal cells. Neuronal cells used in this study were differentiated from human neuroblastoma cell line, SH-SY5Y cells. In vitro ischemia was caused by oxygen-glucose deprivation (OGD) treatment.
Neuronal cell differentiation protocol for SH-SY5Y cells was tested, and results verified with immunocytochemistry using antibodies targeted to mature neuronal cell markers. Differentiated SH- SY5Y cells were predisposed to ASC environment with different culture media and in co-cultural system. Neuronal cell proliferation in co-cultural system was measured using CyQUANT-analysis and neuronal cells were also examined with immunocytochemical assay. Four-hour OGD treatment was conducted to neuronal cell culture and effect was examined using immunocytochemistry with apoptosis marker.
Protocol used for SH-SY5Y neuronal differentiation was successful and cells expressed mature neuronal markers based on immunocytochemistry and fluorescence microscoping. CyQuant analysis showed that SH-SY5Y cells survived well in co-cultural system. OGD treatment time was too short and SH-SY5Y derived neuronal cells remained viable. Also, apoptosis marker was not visible with immunocytochemistry after the OGD treatment.
ASCs ? paracrine effect on neuronal cells after ischemic injury can be studied with human origin SH- SY5Y derived neuronal cell culture as they survived in the co-cultural system. In the present study, too short OGD treatment time prevented examination of the regenerative paracrine effect of ASCs on injured neuronal cells as cells remained intact. Prospective studies could examine different OGD- treatment times to cause ischemic injuries on neuronal cells in order to study if neuroprotectivity is caused by the presence of ASC or the secrete products.
Stroke is one of the leading causes of mortality worldwide. Studies aiming to find new therapeutic methods improving patient’s outcome afterwards are conducted in the field of stem cell research. Adipose stem cells (ASC) may possess neuroprotective ability and they have shown a potential for helping neuronal cells suffered from ischemic injury to regenerate. The aim of the present research is to develop an in vitro ischemia model for studying ASC possible paracrine effect on damaged neuronal cells. Neuronal cells used in this study were differentiated from human neuroblastoma cell line, SH-SY5Y cells. In vitro ischemia was caused by oxygen-glucose deprivation (OGD) treatment.
Neuronal cell differentiation protocol for SH-SY5Y cells was tested, and results verified with immunocytochemistry using antibodies targeted to mature neuronal cell markers. Differentiated SH- SY5Y cells were predisposed to ASC environment with different culture media and in co-cultural system. Neuronal cell proliferation in co-cultural system was measured using CyQUANT-analysis and neuronal cells were also examined with immunocytochemical assay. Four-hour OGD treatment was conducted to neuronal cell culture and effect was examined using immunocytochemistry with apoptosis marker.
Protocol used for SH-SY5Y neuronal differentiation was successful and cells expressed mature neuronal markers based on immunocytochemistry and fluorescence microscoping. CyQuant analysis showed that SH-SY5Y cells survived well in co-cultural system. OGD treatment time was too short and SH-SY5Y derived neuronal cells remained viable. Also, apoptosis marker was not visible with immunocytochemistry after the OGD treatment.
ASCs ? paracrine effect on neuronal cells after ischemic injury can be studied with human origin SH- SY5Y derived neuronal cell culture as they survived in the co-cultural system. In the present study, too short OGD treatment time prevented examination of the regenerative paracrine effect of ASCs on injured neuronal cells as cells remained intact. Prospective studies could examine different OGD- treatment times to cause ischemic injuries on neuronal cells in order to study if neuroprotectivity is caused by the presence of ASC or the secrete products.