Isolation of Viable Follicles from Cryopreserved Human Ovarian Tissue
Introduction: Ovarian tissue freezing is the most effective method to maintain fertility for immature girls and women diagnosed with cancer. Nonetheless, because of the chance that malignant cells might reappearing following tissue transplantation, it is crucial to isolate the follicles from the fro...
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Shahid Sadoughi University of Medical Sciences
2025-01-01
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| Series: | Majallah-i Dānishgāh-i ’Ulūm-i Pizishkī-i Shahīd Ṣadūqī Yazd |
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| Online Access: | http://jssu.ssu.ac.ir/article-1-6261-en.pdf |
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| author | Farnaz Tajbakhsh Somayeh Tavana Mohammad Kazemi Ashtiani Naeimeh Sadat Abtahi Leila Sadat Tahaei Ashraf Moini Rouhollah Fathi |
| author_facet | Farnaz Tajbakhsh Somayeh Tavana Mohammad Kazemi Ashtiani Naeimeh Sadat Abtahi Leila Sadat Tahaei Ashraf Moini Rouhollah Fathi |
| author_sort | Farnaz Tajbakhsh |
| collection | DOAJ |
| description | Introduction: Ovarian tissue freezing is the most effective method to maintain fertility for immature girls and women diagnosed with cancer. Nonetheless, because of the chance that malignant cells might reappearing following tissue transplantation, it is crucial to isolate the follicles from the frozen-thawed ovarian tissue of these individuals and employ them in the process of in vitro maturation process or artificial ovarian framework. This study aimed to assess the application of neutral red (NR) vital dye alongside collagenase IA for effectively isolating viable follicles from the vitrified human ovarian tissue samples.
Methods: Two categories existed: the category with NR and the group without NR. Chopped (0.5×0.5 mm) strips of vitrified-warmed ovarian tissue from 10 transsexual individuals were placed into two falcon tubes with HTCM and Collagenase IA (1mg/ml). Neutral red (NR) was introduced to one of the falcons. Follicles were then isolated mechanically. The morphology, size, and viability of the follicles were assessed. The condition of the follicles was evaluated using fluorescent staining methods involving Calcein-AM and Ethidium homodimer-I. The t-test method was used to evaluate the data.
Results: The number of isolated follicles with Neutral Red (46.50±25/02) exceeded those without NR (6.6±5.58; P < 0/0001). Additionally, according to the morphological studies, a majority of the isolated follicles from the transsexual ovarian cortex were primordial (77.4%), and primary (21.12%) follicles, with only a small number of secondary follicles (1.4%) identified in these tissues. Live/dead staining verified the viability of isolated follicles by displaying a green hue.
Conclusion: The finding indicates that combining collagenase I with the vital dye Neutral red significantly facilitates the of follicles from dense human ovarian tissue. |
| format | Article |
| id | doaj-art-12b56e7bab2b4a9794111b641f86dbc0 |
| institution | Kabale University |
| issn | 2228-5741 2228-5733 |
| language | fas |
| publishDate | 2025-01-01 |
| publisher | Shahid Sadoughi University of Medical Sciences |
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| series | Majallah-i Dānishgāh-i ’Ulūm-i Pizishkī-i Shahīd Ṣadūqī Yazd |
| spelling | doaj-art-12b56e7bab2b4a9794111b641f86dbc02025-01-15T09:29:12ZfasShahid Sadoughi University of Medical SciencesMajallah-i Dānishgāh-i ’Ulūm-i Pizishkī-i Shahīd Ṣadūqī Yazd2228-57412228-57332025-01-01321083308341Isolation of Viable Follicles from Cryopreserved Human Ovarian TissueFarnaz Tajbakhsh0Somayeh Tavana1Mohammad Kazemi Ashtiani2Naeimeh Sadat Abtahi3Leila Sadat Tahaei4Ashraf Moini5Rouhollah Fathi6 Department of Developmental Biology, University of Science and Culture, Tehran, Iran Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran Department of Cell Engineering, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran. Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran Introduction: Ovarian tissue freezing is the most effective method to maintain fertility for immature girls and women diagnosed with cancer. Nonetheless, because of the chance that malignant cells might reappearing following tissue transplantation, it is crucial to isolate the follicles from the frozen-thawed ovarian tissue of these individuals and employ them in the process of in vitro maturation process or artificial ovarian framework. This study aimed to assess the application of neutral red (NR) vital dye alongside collagenase IA for effectively isolating viable follicles from the vitrified human ovarian tissue samples. Methods: Two categories existed: the category with NR and the group without NR. Chopped (0.5×0.5 mm) strips of vitrified-warmed ovarian tissue from 10 transsexual individuals were placed into two falcon tubes with HTCM and Collagenase IA (1mg/ml). Neutral red (NR) was introduced to one of the falcons. Follicles were then isolated mechanically. The morphology, size, and viability of the follicles were assessed. The condition of the follicles was evaluated using fluorescent staining methods involving Calcein-AM and Ethidium homodimer-I. The t-test method was used to evaluate the data. Results: The number of isolated follicles with Neutral Red (46.50±25/02) exceeded those without NR (6.6±5.58; P < 0/0001). Additionally, according to the morphological studies, a majority of the isolated follicles from the transsexual ovarian cortex were primordial (77.4%), and primary (21.12%) follicles, with only a small number of secondary follicles (1.4%) identified in these tissues. Live/dead staining verified the viability of isolated follicles by displaying a green hue. Conclusion: The finding indicates that combining collagenase I with the vital dye Neutral red significantly facilitates the of follicles from dense human ovarian tissue.http://jssu.ssu.ac.ir/article-1-6261-en.pdfhuman follicle isolationneutral redcryopreservationtissue engineeringlive/dead staining. |
| spellingShingle | Farnaz Tajbakhsh Somayeh Tavana Mohammad Kazemi Ashtiani Naeimeh Sadat Abtahi Leila Sadat Tahaei Ashraf Moini Rouhollah Fathi Isolation of Viable Follicles from Cryopreserved Human Ovarian Tissue Majallah-i Dānishgāh-i ’Ulūm-i Pizishkī-i Shahīd Ṣadūqī Yazd human follicle isolation neutral red cryopreservation tissue engineering live/dead staining. |
| title | Isolation of Viable Follicles from Cryopreserved Human Ovarian Tissue |
| title_full | Isolation of Viable Follicles from Cryopreserved Human Ovarian Tissue |
| title_fullStr | Isolation of Viable Follicles from Cryopreserved Human Ovarian Tissue |
| title_full_unstemmed | Isolation of Viable Follicles from Cryopreserved Human Ovarian Tissue |
| title_short | Isolation of Viable Follicles from Cryopreserved Human Ovarian Tissue |
| title_sort | isolation of viable follicles from cryopreserved human ovarian tissue |
| topic | human follicle isolation neutral red cryopreservation tissue engineering live/dead staining. |
| url | http://jssu.ssu.ac.ir/article-1-6261-en.pdf |
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