Amniotic Membrane: Separation of Amniotic Mesoderm from Amniotic Epithelium and Isolation of Their Respective Mesenchymal Stromal and Epithelial Cells

2012 ◽  
Author(s):  
Antonella Barbati ◽  
Maria Grazia Mameli ◽  
Angelo Sidoni ◽  
Gian Carlo Di Renzo
Keyword(s):  
Epithelial Cells ◽  
Amniotic Membrane ◽  
Membrane Separation ◽  
Amniotic Epithelium

386 HORSE AMNION: A SOURCE OF MESENCHYMAL (AMSC) AND EPITHELIAL STEM CELLS

2010 ◽  
Vol 22 (1) ◽  
pp. 349 ◽  
Author(s):  
A. Lange Consiglio ◽  
B. Corradetti ◽  
D. Bizzaro ◽  
M. Cassano ◽  
F. Cremonesi
Keyword(s):  
Stem Cells ◽  
Epithelial Cells ◽  
Amniotic Membrane ◽  
Embryonic Stem ◽  
Specific Pattern ◽  
Limiting Factor ◽  
Epithelial Stem Cells ◽  
Culture Dish ◽  
Isolated Cells ◽  
Differentiation Potential

According to the developmental stage from which they are obtained, stem cells are classified as being embryonic, fetal, or adult. Embryonic stem cells have unlimited self-renewing capacity and multilineage differentiation potential, but the separation of these cells requires destruction of the embryo. Moreover, their clinical application seems to be hindered by the high tumorigenic rate after transplantation. Stem cells derived from adult tissues are considered to be more limited in their potential; although, they are currently the more versatile cells in the clinical field. However, the risk of the immunological rejection of the transplanted stem cells by the recipient is an important limiting factor. In human medicine, stem cells isolated from term placenta are the ideal candidates for disease treatment, specifically because of their plasticity and reduced immunogenicity. The aim of this work was to provide, for the first time, an isolation protocol and the characteristics of the stem cells from horse amniotic membrane, which hold potential uses in equine clinical regenerative medicine. Minimal criteria for stemness definition are adherence to plastic culture dish, formation of fibroblast colony forming units (CFU-F), specific pattern of surface antigen expression, and differentiation potential toward one or more lineages. The amnion is a thin, avascular membrane composed of an epithelial layer and an outer layer of connective tissue. From 3 samples of allantoamnion retrieved at delivery, each amniotic membrane was stripped from the overlying allantois and, for isolation of the epithelial cells, digested with trypsin. After removal of epithelial cells, the AMSC population was obtained by digestion with collagenase and DNase. The cellular yield from term amnion was 10-fold more epithelial cells than AMSC. Isolated cells readily attached to plastic culture dishes. Culture was established in DMEM-HG medium, supplemented with 10% serum and EGF, where the cells proliferated robustly. Epithelial cells displayed typical cuboidal morphology, whereas AMSC were fibroblast-like. Normally, 5 to 6 passages were achieved before proliferation decreased, with a mean of 13.08 and 26.5 cell population doublings after 31 days, respectively, for epithelial cells and AMSC. The mean frequency of CFU-F was, respectively, 1 : 283 and 1:111 for epithelial cells and AMSC. The 2 cellular lines expressed MSC mRNA markers (CD29, CD105, CD44) and were negative for CD34, which was expressed at the fifth passage in both cellular types. Osteogenic differentiation of epithelial stem cells and AMSC was confirmed by von Kossa stain and by an increased expression of osteocalcin and osteopontin. Our preliminary data showed that equine amnion holds apparent potential as a source of presumptive stem cells, which might have widespread clinical applications, but aspects including immunohistochemical study, preclinical experimentation, and immunological properties must be studied.

Embryo co-culture with bovine amniotic membrane stem cells can enhance the cryo-survival of IVF-derived bovine blastocysts comparable with co-culture with bovine oviduct epithelial cells

Zygote ◽  
2020 ◽  
pp. 1-6
Author(s):  
Shayan Nejat-Dehkordi ◽  
Ebrahim Ahmadi ◽  
Abolfazl Shirazi ◽  
Hassan Nazari ◽  
Naser Shams-Esfandabadi
Keyword(s):  
Stem Cells ◽  
Growth Factors ◽  
Epithelial Cells ◽  
Embryonic Development ◽  
Amniotic Membrane ◽  
Biological Activities ◽  
Survival Rates ◽  
Blastocyst Stage ◽  
Oviduct Epithelial Cells

Summary Culture conditions have a profound effect on the quality of in vitro-produced embryos. Co-culturing embryos with somatic cells has some beneficial effects on embryonic development. Considering the ability of stem cells to secrete a broad range of growth factors with different biological activities, we hypothesized that bovine amniotic membrane stem cells (bAMSCs) might be superior to bovine oviduct epithelial cells (BOECs) in supporting embryonic development and enhancing their cryo-survival. Bovine abattoir-derived oocytes were matured and fertilized in vitro. The resultant presumptive zygotes were then cultured up to the blastocyst stage in the following groups: (i) co-culture with bAMSCs, (ii) co-culture with BOECs, and (iii) cell-free culture (Con). Embryos that reached the blastocyst stage were vitrified and warmed, and their post-warming re-expansion, survival and hatching rates were evaluated after 72 h culture. Results showed that the cleavage, blastocyst, and 2 h post-warming re-expansion rates of embryos did not differ between groups. However, their survival rates in BOEC and bAMSC groups were significantly higher compared with the control (72.7, 75.6 and 37.5%, respectively, P < 0.05). In conclusion, our results showed that the cryo-survivability of IVF-derived bovine embryos could be improved through co-culturing with bAMSCs. Moreover, considering the possibility to provide multiple passages from bAMSCs compared with BOECs, due to their stemness properties and their ability to produce growth factors, the use of bAMSCs is a good alternative to BOECs in embryo co-culture systems.

Conjunctival Epithelial Cells Cultured on Human Amniotic Membrane Fail to Transdifferentiate into Corneal Epithelial-Type Cells

Cornea ◽  
1999 ◽  
Vol 18 (2) ◽  
pp. 216 ◽  
Author(s):  
Beom-Jin Cho ◽  
Ali R. Djalilian ◽  
Wesley F. Obritsch ◽  
Dawn M. Matteson ◽  
Chi-Chao Chan ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Amniotic Membrane ◽  
Human Amniotic Membrane ◽  
Corneal Epithelial ◽  
Cells Cultured ◽  
Conjunctival Epithelial Cells

scholarly journals Concise Review: Altered Versus Unaltered Amniotic Membrane as a Substrate for Limbal Epithelial Cells

2018 ◽  
Vol 7 (5) ◽  
pp. 415-427 ◽  
Author(s):  
Tor Paaske Utheim ◽  
Øygunn Aass Utheim ◽  
Panagiotis Salvanos ◽  
Catherine J. Jackson ◽  
Stefan Schrader ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Amniotic Membrane ◽  
Concise Review

Generation of Endostatin by Matrix Metalloproteinase and Cathepsin from Human Limbocorneal Epithelial Cells Cultivated on Amniotic Membrane

2007 ◽  
Vol 48 (2) ◽  
pp. 644 ◽  
Author(s):  
David Hui-Kang Ma ◽  
Jeng-Yuan Yao ◽  
Ming-Tse Kuo ◽  
Lai-Chu See ◽  
Kuei-Ying Lin ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Matrix Metalloproteinase ◽  
Amniotic Membrane

TRANSPLANTATION OF CULTIVATED CORNEAL EPITHELIAL CELLS ON AMNIOTIC MEMBRANE; PRELIMINARY CLINICAL OUTCOMES.

Cornea ◽  
2000 ◽  
Vol 19 (Supplement 2) ◽  
pp. S122
Author(s):  
Jun Shimazaki ◽  
Masayo Aiba ◽  
Shigeto Shimmura ◽  
Kazuo Tsubota
Keyword(s):  
Epithelial Cells ◽  
Clinical Outcomes ◽  
Amniotic Membrane ◽  
Corneal Epithelial Cells ◽  
Corneal Epithelial

In Vitro Culture of Epithelial Cells from Different Anatomical Regions of the Human Amniotic Membrane

10.3791/60551 ◽  
2019 ◽  
Author(s):  
Daniela Avila-González ◽  
Guadalupe García-López ◽  
Néstor E. Díaz-Martínez ◽  
Héctor Flores-Herrera ◽  
Anayansi Molina-Hernández ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
In Vitro Culture ◽  
Amniotic Membrane ◽  
Human Amniotic Membrane
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