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Effect of conditioned medium of umbilical cord-derived mesenchymal stem cells as a culture medium for human granulosa cells: An experimental study
Sumapraja, Kanadi; Hestiantoro, Andon; Liem, Isabella Kurnia; Boediono, Arief & Jacoeb, Teuku Z
Abstract
Background: 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.
Objective: 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.
Materials and Methods: UC-MSCs-CM was collected from umbilical cord stem cell
culture on passage 4. GC 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.
Results: 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).
Conclusion: 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.
Keywords
Conditioned medium; BAX; Survivin; GDF9; IGF-1.
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