Utilization of Human Amniotic Mesenchymal Cells as Feeder Layers to Sustain Propagation of Human Embryonic Stem Cells in the Undifferentiated State

2011 ◽  
Vol 13 (4) ◽  
pp. 281-288 ◽  
Author(s):  
Kehua Zhang ◽  
Zhe Cai ◽  
Yang Li ◽  
Jun Shu ◽  
Lin Pan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Mesenchymal Cells ◽  
Undifferentiated State ◽  
Feeder Layers

100. IN VIVO DIFFERENTIATION OF HUMAN EMBRYONIC STEM CELLS TO UTERINE TISSUE

2009 ◽  
Vol 21 (9) ◽  
pp. 19
Author(s):  
L. Ye ◽  
R. Mayberry ◽  
E. Stanley ◽  
A. Elefanty ◽  
C. Gargett
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Epithelial Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Mesenchymal Cells ◽  
Neonatal Mouse ◽  
Endometrial Tissue ◽  
Human Epithelial Cells

The endometrium undergoes cyclic regeneration. This regeneration has been attributed to adult stem progenitor cells and developmental mechanisms [1, 2]. A better understanding of human endometrial development may shed light on the mechanisms involved in endometrial regeneration and on early origins of adult endometrial disease. The lack of human fetal endometrial tissue has impeded research in early human endometrial development. We hypothesized that directed differentiation of human embryonic stem cells (hESC) to human endometrial tissue by neonatal mouse uterine mesenchyme represents a novel system to study early development of human endometrium. Recent studies have shown that the neonatal mouse uterine mesenchyme is extremely inductive and undergoes reciprocal signalling with human endometrial epithelial cells [3]. Our aim is to establish a xenograft tissue recombination protocol based on a model for human prostate tissue differentiation using hESC [4]. Our method involved formation of embryoid body (EB) with GFP labelled hESC (ENVY) [5] for recombination with 2x0.5mm pieces of epithelial-free uterine mesenchyme from postnatal day 1 mice. Upon fusion in culture, the recombinant tissue is grafted under the kidney capsule of NOD/SCID mice for 4-12 weeks and monitored by in-vivo imaging. Immunohistochemical analysis of recombinant grafts 4 weeks post transplantation (n=4) revealed immature CK8+CK18+Hoxa10+ human epithelial cells surrounded by mouse mesenchymal cells suggesting differentiation of hESC to epithelial cells possibly of endometrial lineage. The ER+PR+SMA+Hoxa10+ mouse mesenchymal cells surrounding human glands differentiated into SMA+ cells possibly via reciprocal signalling from human epithelial cells. At 8 weeks, we found several CK18+/Hoxa10+ human glands co-expressing CA125. These glands are supported by Hoxa10+ human stromal cells. Further experiments are underway to induce the expression of ER and PR in Hoxa10+ epithelial cells which will be crucial in revealing their endometrial lineage.

scholarly journals Human Menstrual Blood-Derived Mesenchymal Cells as New Human Feeder Layer System for Human Embryonic Stem Cells

Cell Medicine ◽  
2014 ◽  
Vol 7 (1) ◽  
pp. 25-35 ◽  
Author(s):  
Danúbia Silva Dos Santos ◽  
Vanessa Carvalho Coelho De Oliveira ◽  
Karina Dutra Asensi ◽  
Leandro Vairo ◽  
Adriana Bastos Carvalho ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Mesenchymal Cells ◽  
Menstrual Blood ◽  
Feeder Layer ◽  
Layer System

scholarly journals Available human feeder cells for the maintenance of human embryonic stem cells

Reproduction ◽  
2004 ◽  
Vol 128 (6) ◽  
pp. 727-735 ◽  
Author(s):  
Jung Bok Lee ◽  
Ji Min Song ◽  
Jeoung Eun Lee ◽  
Jong Hyuk Park ◽  
Sun Jong Kim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Feeder Layer ◽  
Feeder Cells ◽  
Embryonic Fibroblasts ◽  
Differentiation Capacity ◽  
Human Adult ◽  
Feeder Layers

Mouse embryonic fibroblasts (MEFs) have been previously used as feeder cells to support the growth of human embryonic stem cells (hESCs). In this study, human adult uterine endometrial cells (hUECs), human adult breast parenchymal cells (hBPCs) and embryonic fibroblasts (hEFs) were tested as feeder cells for supporting the growth of hESCs to prevent the possibility of contamination from animal feeder cells. Cultured hUECs, hBPCs and hEFs were mitotically inactivated and then plated. hESCs (Miz-hES1, NIH registered) initially established on mouse feeder layers were transferred onto each human feeder layer and split every 5 days. The morphology, expression of specific markers and differentiation capacity of hESCs adapted on each human feeder layer were examined. On hUEC, hBPC and hEF feeder layers, hESCs proliferated for more than 90, 50 and 80 passages respectively. Human feeder-based hESCs were positive for stage-specific embryonic antigen (SSEA)-3 and -4, and Apase; they also showed similar differentiation capacity to MEF-based hESCs, as assessed by the formation of teratomas and expression of tissue-specific markers. However, hESCs cultured on hUEC and hEF feeders were slightly thinner and flatter than MEF- or hBPC-based hESCs. Our results suggest that, like MEF feeder layers, human feeder layers can support the proliferation of hESCs without differentiation. Human feeder cells have the advantage of supporting more passages than when MEFs are used as feeder cells, because hESCs can be uniformly maintained in the undifferentiated stage until they pass through senescence. hESCs established and/or maintained under stable xeno-free culture conditions will be helpful to cell-based therapy.

scholarly journals Induction of fracture repair by mesenchymal cells derived from human embryonic stem cells or bone marrow

2011 ◽  
Vol 29 (12) ◽  
pp. 1804-1811 ◽  
Author(s):  
Anita Undale ◽  
Daniel Fraser ◽  
Theresa Hefferan ◽  
Ross A. Kopher ◽  
James Herrick ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Mesenchymal Cells ◽  
Fracture Repair

scholarly journals Spatio-temporal analyses of OCT4 expression and fate transitions in human embryonic stem cells

2020 ◽  
Author(s):  
Sirio Orozco-Fuentes ◽  
Laura E. Wadkin ◽  
Irina Neganova ◽  
Majlinda Lako ◽  
Rafael A. Barrio ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Computational Models ◽  
Probability Distributions ◽  
Embryonic Stem ◽  
Oct4 Expression ◽  
Undifferentiated State ◽  
Spatio Temporal ◽  
The Times

AbstractOCT4 is one of the transcription factors required to maintain an undifferentiated state in human embryonic stem cells (hESCs). Thus, it is crucial to understand how OCT4 transcription is regulated both at the single-cell and colony level. Here we analyse the changes of OCT4-mCherry intensity expression in hESCs in the presence and absence of the BMP4 morphogenetic protein.We show that OCT4 expression is dynamic, reaching a maximum response 10 h after BMP4 treatment. We obtain the stationary probability distributions that govern the hESCs transitions amongst the different cell states in the presence/absence of BMP4 and establish the times at which the hESCs, that lead to differentiated and pluripotent cells, cluster in the colony. Furthermore, by quantifying the similarities between the OCT4 expression amongst neighbouring hESCs, we show that hESCs express, on average, similar values in their local neighbourhood within the first two days of the experiment and before BMP4 treatment. These results are relevant for the development of mathematical and computational models of adherent hESC colonies.

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