fa تأثیر فاکتور رشد شبه‌انسولین-۱ بر تکثیر سلول‌های بنیادی اسپرماتوگونی بز در شرایط آزمایشگاه عمومى General پژوهشي Research <div style="text-align: justify;"><span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><strong><span lang="FA" style="font-size:9.0pt"><span b="" style="font-family:" zar="">سابقه و هدف:</span></span></strong> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">هدف از انجام مطالعه حاضر ارزیابی تأثیر فاکتور</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">رشد</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">شبه‌انسولینی</span></span><span dir="LTR" style="font-size:9.0pt"><span new="" roman="" style="font-family:" times="">-</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">۱</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">بر تکثیر سلول‌های اسپرماتوگونی بز در محیط آزمایشگاه بود.</span></span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><strong><span lang="AR-SA" style="font-size:9.0pt"><span b="" style="font-family:" zar="">مواد و روش</span></span><span lang="FA" style="font-size:9.0pt"><span b="" style="font-family:" zar="">‌</span></span></strong><span lang="AR-SA" style="font-size:9.0pt"><strong><span b="" style="font-family:" zar="">ها:</span></strong><span b="" lotus="" style="font-family:"><strong> </strong>بیضه</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">بز</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">نابالغ</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">از</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">کشتارگاه گرفته و</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">جداسازی</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">سلول‌های</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">اسپرماتوگونی</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">به</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">روش</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">هضم</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">دومرحله‌ای انجام</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">شد. سلول‌های</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">اسپرماتوگونی</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">در</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">محیط</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">کشت</span></span><span dir="LTR" style="font-size:9.0pt"><span new="" roman="" style="font-family:" times=""> DMEM </span></span><span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">حاوی 5 درصد سرم گوساله جنینی و 1 درصد آنتی‌بیوتیک (پنی</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">‌</span></span><span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">سیلین - استرپتومایسین) کشت</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">داده</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">شد</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">و</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">به گروه شاهد و تیمارهای</span></span> <span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">۱</span></span><span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">،</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">2</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">و</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">3</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">به‌ترتیب</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">مقادیر 0، 10</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">، 40 و 100</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">نانوگرم بر</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">میلی‌لیتر</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">فاکتور</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">رشد</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">شبه</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">‌</span></span><span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">انسولین</span></span><span dir="LTR" style="font-size:9.0pt"><span new="" roman="" style="font-family:" times="">-</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">۱</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">اضافه</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">گردید. داده‌ها</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">با</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">آزمون</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">آنالیز واریانس یک‌طرفه و</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">آزمون</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">تکمیلی</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">توکی</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">در</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">سطح</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">5 درصد آنالیز</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">شدند. جهت</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">شناسایی سلول‌های اسپرماتوگونی</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">از</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">رنگ</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">‌</span></span><span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">آمیزی</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">ایمونوهیستوشیمی</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">علیه</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">آنتی‌ژن</span></span><span dir="LTR" style="font-size:9.0pt"><span new="" roman="" style="font-family:" times=""> PGP9.5 </span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">و همچنین آزمون </span></span><span dir="LTR" style="font-size:9.0pt"><span new="" roman="" style="font-family:" times="">PCR</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">-</span></span> <span dir="LTR" style="font-size:9.0pt"><span new="" roman="" style="font-family:" times="">RT</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">نشانگرهای </span></span><span dir="LTR" style="font-size:9.0pt"><span new="" roman="" style="font-family:" times="">THY-1</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">، </span></span><span dir="LTR" style="font-size:9.0pt"><span new="" roman="" style="font-family:" times="">VASA</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">، </span></span><span dir="LTR" style="font-size:9.0pt"><span new="" roman="" style="font-family:" times="">UCHL-1(PGP9.5)</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">،&nbsp; </span></span><span dir="LTR" style="font-size:9.0pt"><span new="" roman="" style="font-family:" times="">plzf</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:"> و </span></span><span dir="LTR" style="font-size:9.0pt"><span new="" roman="" style="font-family:" times="">OCT-4</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">استفاده</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">شد.</span></span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><span lang="AR-SA" style="font-size:9.0pt"><strong><span b="" style="font-family:" zar="">نتایج:</span></strong><span b="" lotus="" style="font-family:"><strong> </strong>تعداد</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">کلونی‌های</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">سلول‌های</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">اسپرماتوگونی</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">در</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">تیمار</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">3</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">در مقایسه با گروه</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">شاهد</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">و</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">دیگر</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">تیمارها در</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">تمامی روزهای ارزیابی (4،</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">7</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">و</span></span> <span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">10)،</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">به‌طور</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">معنی‌داری بیشتر</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">بود. مساحت</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">کلونی‌های</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">تیمار</span></span> <span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">۱</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">در</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">روزهای</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">7</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">و</span></span> <span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">10</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">کشت،</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">اختلاف</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">معنی‌داری</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">را</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">با تیمار 2 و گروه شاهد نشان</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">داد.</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">همچنین مساحت کلونی‌های تیمار</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">3 در</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">روزهای</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">4،</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">7</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">و</span></span> <span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">10</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">پس</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">از</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">کشت نسبت</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">به</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">گروه</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">شاهد،</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">تیمار</span></span> <span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">۱</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">و</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">تیمار</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">2</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">به‌طور معنی‌داری</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">بیشتر بود. سلول‌های جداشده تمامی نشانگرهای سطحی</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">،</span></span><span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:"> مربوط به سلول‌های بنیادی را بیان کردند.</span></span><span lang="FA" style="font-size:9.0pt"><span style="font-family:"B Lotus""></span></span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span style="text-autospace:none"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><span lang="AR-SA" style="font-size:9.0pt"><strong><span b="" style="font-family:" zar="">نتیجه‌گیری:</span></strong><span b="" lotus="" style="font-family:"> برای کشت کوتاه‌مدت</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">سلول‌های</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">بنیادی</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">اسپرماتوگونی بز</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">در</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">شرایط</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">آزمایشگاه،</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">افزودن</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">فاکتور</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">رشد</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">شبه‌انسولین-</span></span><span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">1 </span></span><span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">با</span></span> <span lang="FA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">دوز</span></span><span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:"> 100</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">نانوگرم</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">بر میلی‌لیتر</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">پیشنهاد</span></span> <span lang="AR-SA" style="font-size:9.0pt"><span b="" lotus="" style="font-family:">می‌شود.</span></span></span></span></span></span></span></span></div> <div style="text-align: justify;"><span style="font-family:Times New Roman;"><span style="color:#000000;"><span style="font-size:12pt"><span style="background:white"><span style="unicode-bidi:embed"><span new="" roman="" times=""><strong>Background:</strong> This study aimed to the effect of insulin-like growth factor-1 on the proliferation of goat spermatogonial stem cells (SSCs) in vitro.</span></span></span></span><br> <span style="font-size:12pt"><span style="background:white"><span style="unicode-bidi:embed"><span new="" roman="" times=""><strong>Materials and Methods: </strong>The testicles of immature goats were obtained from slaughterhouse. Isolation of spermatogonial cells was performed by two-step digestion. The isolated cells were cultured in 4 different groups: 3 groups by adding different doses of IGF-1 (10, 40 and 100 ng/ml) to the basic medium (DMEM containing 5% fetal bovine serum and 1% antibiotic) and the control group (without IGF-1). Data were analyzed by one-way ANOVA and Tukey test at 5% level. Presence of SSCs at the end of culture was determined by reverse transcription polymerase chain reaction (RT-PCR) for several important spermatogonial markers (PGP9.5, THY-1, PLZF, Oct4 and VASA) and immunohistochemical staining against PGP9.5 antigen.</span></span></span></span><br> <span style="font-size:12pt"><span style="background:white"><span style="unicode-bidi:embed"><span new="" roman="" times=""><strong>Results: </strong>In this study, the number of spermatogonial cell colonies in treatment 3 was significantly higher than the control and other treatment groups on days 4, 7, and 10 of culture (<i>P</i><0.05). Also, the surface area of G10 colonies on days 7 and 10 of culture was significantly different (<i>P</i><0.05). The surface area of G100 on days 4, 7, and 10 after culture was significantly higher than control, G10 and G40 (<em>P</em><0.05). The isolated cells expressed all surface markers related to stem cells.</span></span></span></span><br> <span style="font-size:12pt"><span style="background:white"><span style="unicode-bidi:embed"><span new="" roman="" times=""><strong>Conclusion:</strong> According to this results, the addition of 100 ng/ml IGF-1 to culture medium for promoting colonization of goat Spermatogonial stem cells is recommended.</span></span></span></span></span></span></div> فاکتور رشد شبه‌انسولین-۱, سلول بنیادی, اسپرماتوگونی بز, RT-PCR Insulin-like growth factor-1, stem cells, Goat, Spermatogonia, RT-PCR 154 163 http://feyz.kaums.ac.ir/browse.php?a_code=A-10-1499-4&slc_lang=fa&sid=1 Milad Torabi میلاد ترابی milad.torabi137099@gmail.com 2900319475328460041722 2900319475328460041722 No Department of Clinical Sciences, Faculty of Veterinary Medicine, I.R. Iran. گروه علوم درمانگاهی، دانشکده دامپزشکی، دانشگاه رازی، کرمانشاه، ایران Peyman Rahimi-Feyli پیمان رحیمی drp.rahimi@gmail.com 2900319475328460041723 2900319475328460041723 Yes Department of Clinical Sciences, Faculty of Veterinary Medicine, I.R. Iran. گروه علوم درمانگاهی، دانشکده دامپزشکی، دانشگاه رازی، کرمانشاه، ایران Aliasghar Moghaddam علی اصغر مقدم moghaddam@razi.ac.ir 2900319475328460041724 2900319475328460041724 No Department of Clinical Sciences, Faculty of Veterinary Medicine, I.R. Iran. گروه علوم درمانگاهی، دانشکده دامپزشکی، دانشگاه رازی، کرمانشاه، ایران