Matching Items (172)
Description
Current culturing methods allow for human neural progenitor cells to be differentiated into neurons for use in diagnostic tools and disease modeling. An issue arises in the relatively low number of cells that can be successfully expanded and differentiated using these current methods, making the progress of research dependent on these cultures as a large number of cells are needed to conduct relevant assays. This project focuses on the expansion and differentiation of human neural progenitor cells cultured on microcarriers and within a rotating bioreactor system as a way to increase the total number of cells generated. Additionally, cryopreservation and the characteristics of these neurons post thaw is being investigated to create a way for long term storage, as well as, a method for standardizing cell lines between multiple experiments at different time points. The experiments covered in this study are aimed to compare the characteristics of differentiated human neurons, both demented and non-demented cell lines between pre-cryopreservation, freshly differentiated neurons and post-cryopreservation neurons. The assays conducted include immunofluorescence, calcium imaging, quantitative polymerase chain reaction, flow cytometry and ELISA data looking at Alzheimer’s disease traits. With the data collected within this study, the use of bioreactors, in addition to, cryopreservation of human neurons for long term storage can be better implemented into human neural progenitor cell research. Both of these aspects will increase the output of these cultures and potentially remove the bottleneck currently found within human neural disease modeling.
ContributorsHenson, Tanner Jay (Author) / Brafman, David (Thesis director) / Kodibagkar, Vikram (Committee member) / School of Life Sciences (Contributor) / Harrington Bioengineering Program (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
Major Depressive Disorder (MDD) is a common mental disorder that can affect individuals at nearly every stage of life. Women are especially vulnerable to MDD in part, from ovarian hormone level fluctuations. In this thesis, I focused on MDD using a rat model in middle-age to explore potential sex differences in response to a corticosterone (CORT) – induced depressive-like state. Estradiol (E2), a naturally occurring steroid sex hormone in humans and rats, is implicated in mood changes, which is especially prominent during the menopause transition. CORT, a stress hormone, was used to create a depressive-like state in middle-aged female (F) and male (M) rats with their gonads surgically removed. This produced the following independent treatment groups: Sex (F, M), CORT (vehicle = V ml/kg, C 40mg/kg), E2 (V 0.1 ml, E 0.3µg/0.1ml). CORT and E2 injections were injected daily, s.c) for 7 days before behavioral testing began and continued throughout the study when behavior was assessed. For my honor’s thesis, I focused on the social interaction test and elevated plus maze to investigate whether CORT enhanced social avoidance and anxiety, and whether E2 mitigated the CORT effects. In the social interaction test, three new behaviors were assessed (interacting, grooming, and immobility) to better understand exploratory and anxiety profiles of the rats, and these behaviors were quantified over two 5-minute periods in the 10-minute trial. These new quantifications showed that for the female rats, C+E and V+V enhanced the interaction with the novel rat significantly more than an inanimate object, which was not observed in the females given CORT only or E2 only. The males in all conditions showed a significant preference for side with the novel rat compared to the object, however no treatment differences were observed. In both sexes, the overall time spent interacting decreased in the second five minutes of quantification compared to the first five minutes. No effects were observed with grooming or immobility, in part from the high variability across rats. For EPM, female rats treated with CORT and E2 exhibited a lower anxiety index than compared to female rats given CORT only, indicating that E2 mitigated the depressive-like effects of CORT. Males showed no CORT or E2 effects. The result in part supported my hypothesis, as the CORT-treated females exhibited reduced socialization and E2 improved socialization in CORT-treated females, as this was seen in the F-C-E group. Interestingly, CORT failed to produce a depressive-like effect in males in both behavioral tests, which was an unexpected outcome. These results suggest that administration of E2 with CORT mitigated the depressive-like state created by CORT in female rats, however failed to produce these outcomes in males. The outcome of this work will give us insight into the potential mechanisms that may contribute to sex differences with MDD.
ContributorsSladkova, Sara (Author) / Conrad, Cheryl (Thesis director) / Amdam, Gro (Committee member) / Barrett, The Honors College (Contributor) / Department of Psychology (Contributor) / School of Life Sciences (Contributor)
Created2024-05