The Journal of Shahid Sadoughi University of Medical Sciences
مجله علمي پژوهشي دانشگاه علوم پزشكي شهید صدوقی يزد
JSSU
Medical Sciences
http://jssu.ssu.ac.ir
20
journal20
2228-5741
2228-5733
fa
jalali
1403
10
1
gregorian
2025
1
1
32
10
online
1
fulltext
fa
جداسازی فولیکولهای زنده از بافت تخمدان منجمد شده انسان
Isolation of Viable Follicles from Cryopreserved Human Ovarian Tissue
زیست شناسی
Biology
پژوهشي
Original article
<div style="text-align: justify;"><span style="font-size:11pt"><span style="line-height:17.0pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><b><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">مقدمه:</span></span></b> <span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">انجماد بافت تخمدان بهترین روش حفظ باروری برای دختران نابالغ و زنان مبتلا به سرطان است. اما به دلیل احتمال برگشت سلولهای بدخیم بعد از پیوند بافت، جداسازی فولیکول از بافت تخمدان منجمد - ذوبشده این افراد برای بلوغ آزمایشگاهی یا ساخت تخمدان مصنوعی مورد توجه است. هدف از این مطالعه، ارزیابی تأثیر رنگ حیاتی نوترال رد (</span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">Neutral red; NR</span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">) و آنزیم کلاژناز در جداسازی موفق فولیکول زنده از قطعات بافت تخمدان انسانی منجمد شده است.</span></span></span></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:17.0pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><b><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">روش بررسی:</span></span></b><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> در این مطالعه تجربی-</span></span> <span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">مداخلهای، </span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">دو گروه اصلی وجود دارد: گروه حاوی رنگ حیاتی </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">NR</span></span><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> و گروه بدون </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">NR</span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">. قطعات خردشده (0/5×0/5میلیمتر) از بافت تخمدان منجمد شیشهای - ذوبشده ۱۰ فرد تراجنسی، برای هضم آنزیمی به دو فالکون حاوی </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">HTCM</span></span><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> و آنزیم کلاژناز</span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">IA (mg/ml</span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">۱) منتقل شد. به یکی از فالکونها </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">NR</span></span><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> اضافه شد. سپس فولیکولها با روش مکانیکی جداسازی شدند. فولیکولهای جداسازیشده از نظر ریختشناسی و اندازه بررسی شدند. میزان زندهمانی و سلامت فولیکولها با رنگ فلوئورسنت </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">Calcein-AM</span></span><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> و </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">Ethidium homodimer-I</span></span><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> ارزیابی شد. دادهها با روش </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">t-test</span></span><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> توسط </span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"><span style="color:black">نرمافزار </span></span></span><span dir="LTR" new="" roman="" style="font-family:" times=""><span style="color:black">GraphPad Prism </span></span> <span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">مورد ارزیابی قرار گرفت.</span></span></span></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:17.0pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><b><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">نتایج:</span></span></b><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> تعداد فولیکولهای جداسازیشده با </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">NR </span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">(25/02</span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">± </span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">46/50) بیشتر از زمانی است که از </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">NR</span></span><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> برای جداسازی استفاده نشد (۵/58</span></span><span lang="FA" style="font-size:12.0pt"><span new="" roman="" style="font-family:" times="">±</span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> ۶/۶) (۰/0001 > </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">P</span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">). هم چنین با توجه به ارزیابیهای ریختشناسی، بیشتر فولیکولهای جداسازیشده از قشر تخمدان افراد تراجنسی از نوع فولیکولهای بدوی (77/4 درصد) و اولیه (21/12 درصد) بودند و تنها تعداد کمی فولیکولهای ثانویه (1/4 درصد) در بافت تخمدان این افراد وجود داشت. رنگآمیزی </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">Live/dead</span></span><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> زندهبودن فولیکولها را بانمایش رنگ سبز تأیید کرد.</span></span> <span lang="FA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""></span></span></span></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:17.0pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><b><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">نتیجهگیری:</span></span></b> <span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">براساس نتایج استفاده همزمان از کلاژناز </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">IA</span></span><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> و رنگ حیاتی </span></span><span dir="LTR" style="font-size:10.0pt"><span new="" roman="" style="font-family:" times="">Neutral red</span></span><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> نقش مهمی در سهولت جداسازی فولیکول از بافت تخمدان متراکم انسان دارد.</span></span></span></span></span></span></span><br>
</div>
<div style="text-align: justify;"><span style="font-size:11pt"><span style="line-height:14.0pt"><span calibri="" style="font-family:"><b><span new="" roman="" style="font-family:" times="">Introduction:</span></b> <span new="" roman="" style="font-family:" times="">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</span><span new="" roman="" style="font-family:" times="">.</span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:14.0pt"><span calibri="" style="font-family:"><b><span new="" roman="" style="font-family:" times="">Methods:</span></b> <span lang="EN" new="" roman="" style="font-family:" times="">Two categories existed: the category with NR and the group without NR. </span><span new="" roman="" style="font-family:" times="">C</span><span lang="EN" new="" roman="" style="font-family:" times="">hopped (0.5×0.5 mm) strips of vitrified-warmed ovarian tissue from 10 transsexual individuals were placed into two falcon tubes with </span><span new="" roman="" style="font-family:" times="">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 </span><span new="" roman="" style="font-family:" times=""><span style="color:black">Calcein-AM and Ethidium homodimer-I. </span></span><span lang="EN" new="" roman="" style="font-family:" times=""><span style="color:black">The t-test method was used to evaluate the data.</span></span><span style="font-family:"Times New Roman",serif"></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:14.0pt"><span calibri="" style="font-family:"><b><span new="" roman="" style="font-family:" times="">Results: </span></b><span new="" roman="" style="font-family:" times="">The number of </span><span new="" roman="" style="font-family:" times="">isolated follicles with Neutral Red (46.50±25/02) exceeded those without NR (6.6±5.58; <i>P</i> < 0/0001).<span style="color:black"> 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.</span></span><span style="font-family:"Times New Roman",serif"></span></span></span></span><br>
<span style="font-size:11pt"><span style="line-height:14.0pt"><span calibri="" style="font-family:"><b><span new="" roman="" style="font-family:" times="">Conclusion: </span></b><span new="" roman="" style="font-family:" times="">The finding indicates that combining collagenase I with the vital dye Neutral red significantly facilitates the of follicles from dense human ovarian tissue.</span></span></span></span><br>
</div>
جداسازی فولیکول انسانی, Neutral Red, انجماد, مهندسی بافت, رنگآمیزی Live/dead
human follicle isolation, Neutral Red, cryopreservation, tissue engineering, live/dead staining.
8330
8341
http://jssu.ssu.ac.ir/browse.php?a_code=A-10-1556-2&slc_lang=fa&sid=1
Farnaz
Tajbakhsh
فرناز
تاج بخش
tajbakhsh_farnaz@yahoo.com