- 作者: Kavitha Sivasubramaiyan; Swapnil Totey; Vijay Bhat; Satish M Totey; Kaushik Deb
- 作者服務機構: Stempeutics Research Pvt Ltd., Bangalore, India
- 中文摘要: --
- 英文摘要:
Trophoblast differentiation and formation of the placenta are important events linked to postimplantation
embryonic development. Models mimicking the biology of trophoblast differentiation
in a post-implantation maternal microenvironment are needed for understanding disorders like
placental-ischemia or for applications in drug-screening, and would help in overcoming the ethical
impasse on using human embryos for such research. Here we attempt to create such a model by
using embryoid bodies (EBs) and a biomimetic platform composed of a bilayer of fibronectin and
gelatin on top of low-melting agarose. Using this model we test the hypothesis that cystic-EBs (day
30) that resemble blastocysts morphologically, are better sources as compared to noncytic EBs
(day 10), for functional trophoblast differentiation; and that the Rho kinases inhibitor Y27632 can
enhance this differentiation. Non/cytic EBs with/out Y27632 were grown on this platform for 28
days, and screened from secretion and expression of trophoblast and other lineage markers using
ECLIA, RT-PCR, and Immunofluorescence. All EBs attached on this surface and rapidly proliferated
into hCG and progesterone (P2) secreting functional trophoblast cells. However, the cells derived
from cytic-EBs and cytic-EBs+ Y27632 showed the maximum secretion of these hormones and
expressed IGF2, supporting our hypothesis. Also Y27632 reduced extraembryonic endoderm and
trophoblast lineage differentiation from early noncystic-EBs, whereas, it specifically enhanced the
induction of trophoblast and multinucleated syncitiotrophoblast differentiation from late cystic-
EBs. In vivo trophoblast differentiation can be replicated in fibronectin based biomaterials, using
cytic-EBs and by maneuvering the Rho-ROCK pathways. Response of EBs to a compound may vary
temporally, and determination of their right stage is crucial for applications in directeddifferentiation
or drug-screening. - 中文關鍵字: --
- 英文關鍵字: --