International Journal of Reproductive BioMedicine
International Journal of Reproductive BioMedicine
IJRM
Medical Sciences
http://ijrm.ir
1
admin
2476-4108
2476-3772
10.29252/ijrm
en
jalali
1400
9
1
gregorian
2021
12
1
19
12
online
1
fulltext
en
تأثیر محیط شرطی شده سلول های بنیادی مزانشیمی مشتق از بند ناف به عنوان محیط کشت برای سلول های گرانولوزای انسانی: یک مطالعه تجربی
Effect of conditioned medium of umbilical cord-derived mesenchymal stem cells as a culture medium for human granulosa cells: An experimental study
Reproductive Endocrinology
Original Article
Original Article
<div style="text-align: justify;"><strong><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">مقدمه: </span></span></strong><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">محیط شرطی شده سلول­های بنیادی مزانشیمی مشتق از بند ناف (</span></span><span dir="LTR"><span style="font-size:10.0pt;">UC-MSCs-CM</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">) ترشحاتی با خواص ضد آپوپتوتیک تولید می</span></span><span dir="LTR"><span style="font-family:Arial,sans-serif;"><span style="font-size:11.0pt;">­</span></span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">کند و این پتانسیل را دارد که از آپوپتوز سلول</span></span><span dir="LTR"><span style="font-family:Arial,sans-serif;"><span style="font-size:11.0pt;">­</span></span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">های گرانولوزا (</span></span><span dir="LTR"><span style="font-size:10.0pt;">GC</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">) در طی تحریک کنترل شده تخمدان جلوگیری کند.</span></span><strong><span dir="LTR"><span style="font-family:Arial,sans-serif;"><span style="font-size:11.0pt;"></span></span></span></strong><br>
<strong><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">هدف: </span></span></strong><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">مشاهده اثر </span></span><span dir="LTR"><span style="font-size:10.0pt;">UC-MSCs-CM</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;"> بر تعامل بین پروتئین</span></span><span dir="LTR"><span style="font-family:Arial,sans-serif;"><span style="font-size:11.0pt;">­</span></span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">های پرو و ضد آپوپتوز و تأثیر تولید فاکتور تمایز رشد 9 (</span></span><span dir="LTR"><span style="font-size:10.0pt;">GDF9</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">) در </span></span><span dir="LTR"><span style="font-size:10.0pt;">GC</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">بود.</span></span><br>
<strong><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">مواد و روش ­ها: </span></span></strong><span dir="LTR"><span style="font-size:10.0pt;">UC-MSCs-CM</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;"> از کشت سلولهای بنیادی بند ناف در پاساژ 4 جمعآوری شد. </span></span><span dir="LTR"><span style="font-size:10.0pt;">GCs</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">از 23 زن که تحت لقاح آزمایشگاهی قرار گرفته بودند، کشت داده شدند و به مدت 24 ساعت در معرض </span></span><span dir="LTR"><span style="font-size:10.0pt;">UC-MSCs-CM</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;"> قرار گرفتند. سپس </span></span><span dir="LTR"><span style="font-size:10.0pt;">RNA GC</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;"> استخراج شد و بیان </span></span><span dir="LTR"><span style="font-size:10.0pt;">mRNA</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">مربوط به </span></span><span dir="LTR"><span style="font-size:10.0pt;">BCL-2 X (BAX)</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">، </span></span><span dir="LTR"><span style="font-size:10.0pt;">survivin</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">و </span></span><span dir="LTR"><span style="font-size:10.0pt;">GDF9</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">با استفاده از </span></span><span dir="LTR"><span style="font-size:10.0pt;">Real-time PCR</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">کمی آنالیز شد. محیط کشت مصرف شده </span></span><span dir="LTR"><span style="font-size:10.0pt;">GC</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">برای اندازه</span></span><span dir="LTR"><span style="font-family:Arial,sans-serif;"><span style="font-size:11.0pt;">­</span></span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">گیری فاکتور رشد انسولین 1 با استفاده از روش الایزا، جمع</span></span><span dir="LTR"><span style="font-family:Arial,sans-serif;"><span style="font-size:11.0pt;">­</span></span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">آوری شد. </span></span><br>
<strong><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">نتایج: </span></span></strong><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">بیان </span></span><span dir="LTR"><span style="font-size:10.0pt;">BAX</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">پس از قرار گرفتن در معرض </span></span><span dir="LTR"><span style="font-size:10.0pt;">UC-MSCs-CM</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;"> به طور قابل توجهی متفاوت بود (</span></span><span dir="LTR"><span style="font-size:10.0pt;">4.09E-7</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;"> در مقابل </span></span><span dir="LTR"><span style="font-size:10.0pt;">3.74E-7</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">، 02/</span></span><span style="font-family:B Nazanin;"><span style="font-size:11.0pt;">0 </span></span><span style="font-size:10.0pt;">=</span> <span dir="LTR"><span style="font-size:10.0pt;">p</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">). پس از قرار گرفتن در معرض </span></span><span dir="LTR"><span style="font-size:10.0pt;">UC-MSCs-CM</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;"> هیچ تغییر قابل توجهی در </span></span><span dir="LTR"><span style="font-size:10.0pt;">survivin</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">، نسبت </span></span><span dir="LTR"><span style="font-size:10.0pt;">BAX/survivin</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">و بیان </span></span><span dir="LTR"><span style="font-size:10.0pt;">GDF9</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">رخ نداد (05/</span></span><span style="font-family:B Nazanin;"><span style="font-size:11.0pt;">0</span></span><span style="font-size:10.0pt;"> < </span><span dir="LTR"><span style="font-size:10.0pt;">p</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">). سطح </span></span><span dir="LTR"><span style="font-size:10.0pt;">IGF-1 CM</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">پس از استفاده از </span></span><span dir="LTR"><span style="font-size:10.0pt;">CM</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">به عنوان محیط کشت برای </span></span><span dir="LTR"><span style="font-size:10.0pt;">GC</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">به طور قابل توجهی بالاتر بود (28/2 در مقابل 72/1</span></span> <span style="font-size:11.0pt;">±</span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;"> 07/3، 001/</span></span><span style="font-family:B Nazanin;"><span style="font-size:11.0pt;">0 </span></span><span style="font-size:10.0pt;">≥</span><span dir="LTR"><span style="font-size:10.0pt;">p </span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">). همبستگی مثبت و معناداری بین </span></span><span dir="LTR"><span style="font-size:10.0pt;">Survivin</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">و </span></span><span dir="LTR"><span style="font-size:10.0pt;">GDF9</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">یافت شد (</span></span><span style="font-family:B Nazanin;"><span style="font-size:11.0pt;">966/0 </span></span><span style="font-size:10.0pt;">= </span><span dir="LTR"><span style="font-size:10.0pt;">r</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">، 001/</span></span><span style="font-family:B Nazanin;"><span style="font-size:11.0pt;">0 </span></span><span style="font-size:10.0pt;">≥ </span><span dir="LTR"><span style="font-size:10.0pt;">p</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">).</span></span><br>
<strong><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">نتیجه ­گیری: </span></span></strong><span dir="LTR"><span style="font-size:10.0pt;">IGF-1</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;"> تولید شده توسط </span></span><span dir="LTR"><span style="font-size:10.0pt;">UC-MSCs-CM</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">میتواند به روش پاراکرین از طریق گیرنده </span></span><span dir="LTR"><span style="font-size:10.0pt;">IGF</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">کار کند که میتواند </span></span><span dir="LTR"><span style="font-size:10.0pt;">BAX</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">را مهار کرده و تولید </span></span><span dir="LTR"><span style="font-size:10.0pt;">GDF9</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">را حفظ کند. علاوه بر این، تحت تأثیر </span></span><span dir="LTR"><span style="font-size:10.0pt;">UC-MSCs-CM</span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">، </span></span><span dir="LTR"><span style="font-size:10.0pt;">GC</span></span><span style="font-family:B Nazanin;"><span style="font-size:11.0pt;">ها</span></span><span style="font-family:B Nazanin;"><span style="font-size:11.0pt;"> همچنین </span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">قادر به تولید </span></span><span dir="LTR"><span style="font-size:10.0pt;">IGF-1</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">هستند که می</span></span><span dir="LTR"><span style="font-family:Arial,sans-serif;"><span style="font-size:11.0pt;">­</span></span></span><span style="font-family:B Mitra;"><span style="font-size:11.0pt;">تواند </span></span><span dir="LTR"><span style="font-size:10.0pt;">GC</span></span> <span style="font-family:B Mitra;"><span style="font-size:11.0pt;">را از طریق فرآیندهای اتوکرین تحت تأثیر قرار دهد.</span></span><strong><span dir="LTR"><span style="font-family:Arial,sans-serif;"><span style="font-size:11.0pt;"></span></span></span></strong><br>
</div>
<p style="text-align: justify;"><strong>Background</strong><strong>:</strong> The umbilical cord-derived mesenchymal stem cells conditioned medium (UC-MSCs-CM) produces secretomes with anti-apoptotic properties, and has the potential to prevent apoptosis of granulosa cells (GC) during controlled ovarian hyperstimulation.<br>
<strong>Objective:</strong> To observe the effect of UC-MSCs-CM on the interaction between pro- and anti-apoptotic proteins and the influence of growth differentiation factor 9 (GDF9) production in GC.<br>
<strong>Materials and Methods: </strong>UC-MSCs-CM was collected from umbilical cord stem cell culture on passage 4. GCs from 23 women who underwent in vitro fertilization were cultured and exposed to UC-MSCs-CM for 24 hr. Then RNA of the GC was extracted and the mRNA expression of BCL-2 associated X (BAX), survivin and GDF9 were analysed using quantitative real-time PCR. The spent culture media of the GC were collected for measurement of insulin growth factor 1 using ELISA.<br>
<strong>Results:</strong> The expression of BAX was significantly different after UC-MSCs-CM exposure (4.09E-7 vs. 3.74E-7, p = 0.02). No significant changes occurred in survivin, BAX/survivin ratio, and GDF9 expression after UC-MSCs-CM exposure (p > 0.05). The IGF-1 level of the CM was significantly higher after the CM was used as a culture medium for GC (2.28 vs. 3.07 ± 1.72, p ≤ 0.001). A significant positive correlation was found between survivin and GDF9 (r = 0.966, p ≤ 0.001).<br>
<strong>Conclusion:</strong> IGF-1 produced by UC-MSCs-CM can work in paracrine fashion through the IGF receptor, which can inhibit BAX and maintain GDF9 production. Moreover, under the influence of UC-MSCs-CM, GC are also capable of producing IGF-1, which can impact GC through autocrine processes.</p>
محیط شرطی, BAX, Survivin, GDF9, IGF-1.
Conditioned medium, BAX, Survivin, GDF9, IGF-1.
1037
1044
http://ijrm.ir/browse.php?a_code=A-10-1203-1&slc_lang=en&sid=1
Kanadi
Sumapraja
kanadisuma@yahoo.com