65 Testing the effects of rigid encapsulations on bovine primordial follicle quiescence versus growth
OBJECTIVES/GOALS: There is an interest in developing a bioprosthetic ovary for ovarian tissue transplantation. The properties of the ovarian extracellular matrix need to be better understood in order to replicate the human ovary. We tested the effects of an encapsulating hydrogel at different rigidi...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Cambridge University Press
2024-04-01
|
| Series: | Journal of Clinical and Translational Science |
| Online Access: | https://www.cambridge.org/core/product/identifier/S2059866124000712/type/journal_article |
| _version_ | 1797226949892898816 |
|---|---|
| author | Kathryn L McElhinney Erin E Rowell Monica M Laronda |
| author_facet | Kathryn L McElhinney Erin E Rowell Monica M Laronda |
| author_sort | Kathryn L McElhinney |
| collection | DOAJ |
| description | OBJECTIVES/GOALS: There is an interest in developing a bioprosthetic ovary for ovarian tissue transplantation. The properties of the ovarian extracellular matrix need to be better understood in order to replicate the human ovary. We tested the effects of an encapsulating hydrogel at different rigidities on bovine primordial follicle activation, growth, and survival. METHODS/STUDY POPULATION: Bovine primordial follicles were isolated from ovarian cortex. A mean of 9.9 follicles (range 3-24) were encapsulated per bead in either 1% or 5 % alginate across 4 experiments. The encapsulated follicles were subsequently crosslinked in a calcium sulfate solution. Follicles were then cultured for 8 days with light microscopy imaging taken every other day along with media exchanges. Follicles were then examined using immunofluorescence. Growth and survival curves were constructed and all statistical analyses were performed using Graph Pad Prism 9. RESULTS/ANTICIPATED RESULTS: A total of 372 follicles were encapsulated across 32 beads (16 in 1% alginate and 16 in 5% alginate). There were no differences in initial follicle size between the two conditions (33.53 µm vs. 32.45 µm, p=0.47). At the end of 8 days, there was no difference between follicle size (59.55 vs. 56.06, p=0.48). Additionally, there was no difference in survival between 1% and 5% alginate encapsulation (57.75% vs. 52.43%. p=0.40). Immunofluorescence is being performed on encapsulated follicles to confirm the presence of DDX4, a molecular marker of oocytes, after 8 days in culture. Additional encapsulated follicles have also been submitted for histologic sectioning and hematoxylin and eosin staining to better characterize the viability and health of these follicles after 8 days in culture. DISCUSSION/SIGNIFICANCE: There was no significant difference in growth or survival between primordial follicles cultured in 1% or 5% alginate gels. Immunofluorescent analysis confirmed the presence of viable follicles at the end of 8 days of culture. Future work needs to further explore how factors in the ovarian extracellular matrix impact follicle maintenance and growth. |
| first_indexed | 2024-04-24T14:33:02Z |
| format | Article |
| id | doaj.art-32dde9f12dc84e43b0741356cec4e691 |
| institution | Directory Open Access Journal |
| issn | 2059-8661 |
| language | English |
| last_indexed | 2024-04-24T14:33:02Z |
| publishDate | 2024-04-01 |
| publisher | Cambridge University Press |
| record_format | Article |
| series | Journal of Clinical and Translational Science |
| spelling | doaj.art-32dde9f12dc84e43b0741356cec4e6912024-04-03T02:00:00ZengCambridge University PressJournal of Clinical and Translational Science2059-86612024-04-018171810.1017/cts.2024.7165 Testing the effects of rigid encapsulations on bovine primordial follicle quiescence versus growthKathryn L McElhinney0Erin E Rowell1Monica M Laronda2Ann & Robert H. Lurie Children’s Hospital of Chicago, Northwestern University Feinberg School of MedicineAnn & Robert H. Lurie Children’s Hospital of Chicago, Northwestern University Feinberg School of MedicineAnn & Robert H. Lurie Children’s Hospital of Chicago, Northwestern University Feinberg School of MedicineOBJECTIVES/GOALS: There is an interest in developing a bioprosthetic ovary for ovarian tissue transplantation. The properties of the ovarian extracellular matrix need to be better understood in order to replicate the human ovary. We tested the effects of an encapsulating hydrogel at different rigidities on bovine primordial follicle activation, growth, and survival. METHODS/STUDY POPULATION: Bovine primordial follicles were isolated from ovarian cortex. A mean of 9.9 follicles (range 3-24) were encapsulated per bead in either 1% or 5 % alginate across 4 experiments. The encapsulated follicles were subsequently crosslinked in a calcium sulfate solution. Follicles were then cultured for 8 days with light microscopy imaging taken every other day along with media exchanges. Follicles were then examined using immunofluorescence. Growth and survival curves were constructed and all statistical analyses were performed using Graph Pad Prism 9. RESULTS/ANTICIPATED RESULTS: A total of 372 follicles were encapsulated across 32 beads (16 in 1% alginate and 16 in 5% alginate). There were no differences in initial follicle size between the two conditions (33.53 µm vs. 32.45 µm, p=0.47). At the end of 8 days, there was no difference between follicle size (59.55 vs. 56.06, p=0.48). Additionally, there was no difference in survival between 1% and 5% alginate encapsulation (57.75% vs. 52.43%. p=0.40). Immunofluorescence is being performed on encapsulated follicles to confirm the presence of DDX4, a molecular marker of oocytes, after 8 days in culture. Additional encapsulated follicles have also been submitted for histologic sectioning and hematoxylin and eosin staining to better characterize the viability and health of these follicles after 8 days in culture. DISCUSSION/SIGNIFICANCE: There was no significant difference in growth or survival between primordial follicles cultured in 1% or 5% alginate gels. Immunofluorescent analysis confirmed the presence of viable follicles at the end of 8 days of culture. Future work needs to further explore how factors in the ovarian extracellular matrix impact follicle maintenance and growth.https://www.cambridge.org/core/product/identifier/S2059866124000712/type/journal_article |
| spellingShingle | Kathryn L McElhinney Erin E Rowell Monica M Laronda 65 Testing the effects of rigid encapsulations on bovine primordial follicle quiescence versus growth Journal of Clinical and Translational Science |
| title | 65 Testing the effects of rigid encapsulations on bovine primordial follicle quiescence versus growth |
| title_full | 65 Testing the effects of rigid encapsulations on bovine primordial follicle quiescence versus growth |
| title_fullStr | 65 Testing the effects of rigid encapsulations on bovine primordial follicle quiescence versus growth |
| title_full_unstemmed | 65 Testing the effects of rigid encapsulations on bovine primordial follicle quiescence versus growth |
| title_short | 65 Testing the effects of rigid encapsulations on bovine primordial follicle quiescence versus growth |
| title_sort | 65 testing the effects of rigid encapsulations on bovine primordial follicle quiescence versus growth |
| url | https://www.cambridge.org/core/product/identifier/S2059866124000712/type/journal_article |
| work_keys_str_mv | AT kathrynlmcelhinney 65testingtheeffectsofrigidencapsulationsonbovineprimordialfolliclequiescenceversusgrowth AT erinerowell 65testingtheeffectsofrigidencapsulationsonbovineprimordialfolliclequiescenceversusgrowth AT monicamlaronda 65testingtheeffectsofrigidencapsulationsonbovineprimordialfolliclequiescenceversusgrowth |