Isolation and Differentiation of Amniotic Membrane Stem Cells Into Keratinocytes

Author:

Lan Dam Thi Phuong12ORCID,Binh Pham Thai32,Giang Nguyen Thi Quynh3,Van Mao Can4,Chung Dang Thanh4,Van Diep Nong5,Trung Do Minh6,Van Tran Pham1ORCID

Affiliation:

1. Department of Biochemistry, Military Hospital 103, Vietnam Military Medical University, Hanoi, Vietnam

2. * Both the authors share the first authorship

3. National Hospital of Endocrinology, Hanoi, Vietnam

4. Physiology Department, Vietnam Military Medical University, Hanoi, Vietnam

5. Department of Biochemistry, Backan Hospital, Backan, Vietnam

6. Research Institute of Medicine & Pharmacy, Vietnam Military Medical University, Hanoi, Vietnam

Abstract

The human amniotic membrane is a highly abundant and readily available tissue that may be useful for regenerative medicine and cell therapy. The amniotic membrane stem cells can differentiate into multiple cell lineages; they have low immunogenicity and anti-inflammatory functions. This research aims to examine the protocols for the isolation of human amniotic membrane stem cells, including their phenotypic characterization and in vitro potential for differentiation toward keratinocytes. Human placentas were obtained from selected cesarean-sectioned births. We isolated amniotic stem cells by trypsin and collagenase B digestion and centrifuged with Percoll. After monolayer expansion of adherent cells, the cells were characterized by immunocytology with octamer-binding transcription factor 4 and differentiated into keratinocytes by treating the cells with insulin, hydrocortisone, BMP-4, and vitamin C. Protocol for isolation of stem cells from amniotic membrane has high efficiency. Differentiation markers of stem cells into keratinocytes, such as vimentin, cytokeratin (CK) 14, and CK19, were determined by reverse transcription-polymerase chain reaction increase over time in culture. Stem cells isolated from the amniotic membrane can differentiate into keratinocytes. It has opened the prospect of using stem cells to regenerate skin and clinical applications.

Funder

Ministry of science and technology, Vietnam

Publisher

SAGE Publications

Subject

Transplantation,Cell Biology,Biomedical Engineering

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