Large-scale Isolation, Expansion and Characterization of Human Amniotic Epithelial Cells

Overview

Journal Int J Stem Cells

Date 2018 May 30

PMID 29843193

Citations 9

Authors

Sanjay Gottipamula

K N Sridhar

Affiliations

Soon will be listed here.

Abstract

Background And Objectives: The human Amniotic epithelial cells (AME) derived from amniotic membrane of placenta have been considered as the potential fetal stem cell source with minimal or no ethical concerns and are important therapeutic tool for anti-fibrotic and regenerative therapies.

Methods And Results: Here, we evaluated the isolation, media screening, scale-up and characterization of AME cells. The isolation, expansion of AMEs were performed by sequential passaging and growth kinetics studies. The AMEs were characterized using immunocytochemistry, immunophenotyping, differentiation, and anti-fibrotic assays. The growth kinetics study revealed that the AME cultured in Ultraculture (UC) and DMEM knockout (DMEM-KO) have prominently higher growth rate compared to others. Overall, the AMEs cultured from 5 different media retained basic morphological characteristics and the functional characteristics.

Conclusions: Our result suggests that the AMEs can be successfully cultured in UC based complete media without losing its epithelial cell characteristics even after passaging for passage 2 (P2). However, a careful and methodical pre-clinical and clinical translation studies need to be conducted to show its safety and efficacy.

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References

Miki T, Lehmann T, Cai H, Stolz D, Strom S . Stem cell characteristics of amniotic epithelial cells. Stem Cells. 2005; 23(10):1549-59. DOI: 10.1634/stemcells.2004-0357. View

You H, Han S . Cell therapy for wound healing. J Korean Med Sci. 2014; 29(3):311-9. PMC: 3945123. DOI: 10.3346/jkms.2014.29.3.311. View

Varkey M, Ding J, Tredget E . Advances in Skin Substitutes-Potential of Tissue Engineered Skin for Facilitating Anti-Fibrotic Healing. J Funct Biomater. 2015; 6(3):547-63. PMC: 4598670. DOI: 10.3390/jfb6030547. View

Mason C, Brindley D, Culme-Seymour E, Davie N . Cell therapy industry: billion dollar global business with unlimited potential. Regen Med. 2011; 6(3):265-72. DOI: 10.2217/rme.11.28. View

Miki T, Marongiu F, Dorko K, Ellis E, Strom S . Isolation of amniotic epithelial stem cells. Curr Protoc Stem Cell Biol. 2010; Chapter 1:Unit 1E.3. DOI: 10.1002/9780470151808.sc01e03s12. View

Jiawen S, Jianjun Z, Jiewen D, Dedong Y, Hongbo Y, Jun S . Osteogenic differentiation of human amniotic epithelial cells and its application in alveolar defect restoration. Stem Cells Transl Med. 2014; 3(12):1504-13. PMC: 4250213. DOI: 10.5966/sctm.2014-0118. View

Ochsenbein-Kolble N, Bilic G, Hall H, Huch R, Zimmermann R . Inducing proliferation of human amnion epithelial and mesenchymal cells for prospective engineering of membrane repair. J Perinat Med. 2003; 31(4):287-94. DOI: 10.1515/JPM.2003.040. View

Di Germanio C, Bernier M, de Cabo R, Barboni B . Amniotic Epithelial Cells: A New Tool to Combat Aging and Age-Related Diseases?. Front Cell Dev Biol. 2016; 4:135. PMC: 5118838. DOI: 10.3389/fcell.2016.00135. View

Matikainen T, Laine J . Placenta--an alternative source of stem cells. Toxicol Appl Pharmacol. 2005; 207(2 Suppl):544-9. DOI: 10.1016/j.taap.2005.01.039. View

10.

Ilancheran S, Michalska A, Peh G, Wallace E, Pera M, Manuelpillai U . Stem cells derived from human fetal membranes display multilineage differentiation potential. Biol Reprod. 2007; 77(3):577-88. DOI: 10.1095/biolreprod.106.055244. View

11.

Silini A, Cargnoni A, Magatti M, Pianta S, Parolini O . The Long Path of Human Placenta, and Its Derivatives, in Regenerative Medicine. Front Bioeng Biotechnol. 2015; 3:162. PMC: 4609884. DOI: 10.3389/fbioe.2015.00162. View

12.

Wu Z, Hui G, Lu Y, Liu T, Huang Q, Guo L . Human amniotic epithelial cells express specific markers of nerve cells and migrate along the nerve fibers in the corpus callosum. Neural Regen Res. 2015; 7(1):41-5. PMC: 4354114. DOI: 10.3969/j.issn.1673-5374.2012.01.007. View

13.

Diaz-Prado S, Muinos-Lopez E, Hermida-Gomez T, Rendal-Vazquez M, Fuentes-Boquete I, de Toro F . Multilineage differentiation potential of cells isolated from the human amniotic membrane. J Cell Biochem. 2010; 111(4):846-57. DOI: 10.1002/jcb.22769. View

14.

Saito S, Yokoyama K, Tamagawa T, Ishiwata I . Derivation and induction of the differentiation of animal ES cells as well as human pluripotent stem cells derived from fetal membrane. Hum Cell. 2006; 18(3):135-41. DOI: 10.1111/j.1749-0774.2005.tb00003.x. View

15.

Manuelpillai U, Lourensz D, Vaghjiani V, Tchongue J, Lacey D, Tee J . Human amniotic epithelial cell transplantation induces markers of alternative macrophage activation and reduces established hepatic fibrosis. PLoS One. 2012; 7(6):e38631. PMC: 3375296. DOI: 10.1371/journal.pone.0038631. View

16.

McDonald C, Payne N, Sun G, Moussa L, Siatskas C, Lim R . Immunosuppressive potential of human amnion epithelial cells in the treatment of experimental autoimmune encephalomyelitis. J Neuroinflammation. 2015; 12:112. PMC: 4457975. DOI: 10.1186/s12974-015-0322-8. View

17.

Ackermann K, Borgia S, Korting H, Mewes K, Schafer-Korting M . The Phenion full-thickness skin model for percutaneous absorption testing. Skin Pharmacol Physiol. 2009; 23(2):105-12. DOI: 10.1159/000265681. View

18.

Parolini O, Alviano F, Bagnara G, Bilic G, Buhring H, Evangelista M . Concise review: isolation and characterization of cells from human term placenta: outcome of the first international Workshop on Placenta Derived Stem Cells. Stem Cells. 2007; 26(2):300-11. DOI: 10.1634/stemcells.2007-0594. View

19.

Yeager A, Singer H, Buck J, Matalon R, Brennan S, OToole S . A therapeutic trial of amniotic epithelial cell implantation in patients with lysosomal storage diseases. Am J Med Genet. 1985; 22(2):347-55. DOI: 10.1002/ajmg.1320220219. View

20.

Vig K, Chaudhari A, Tripathi S, Dixit S, Sahu R, Pillai S . Advances in Skin Regeneration Using Tissue Engineering. Int J Mol Sci. 2017; 18(4). PMC: 5412373. DOI: 10.3390/ijms18040789. View