ISOLATION AND IMMUNOPHENOTYPJNG OF HUMAN AMNIOTIC EPITHELIAL CELLS

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TABATABAEI MERAJ; ZARNANI AMIR HOSSEIN; NIKOO SHOHREH; KESHAVARZI BAHAREH; RAMZANI TEHRANI FAHIMEH; AREFI SOHEILA; MIRZADEGAN EBRAHIM; MOSAFFA NARIMAN

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Journal: Research in Medicine Year:2013 | Volume:37 | Issue:1 Page(s): 1-7

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Title

ISOLATION AND IMMUNOPHENOTYPJNG OF HUMAN AMNIOTIC EPITHELIAL CELLS

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Keywords

PLACENTAQ3

STEM CELLQ2

AMNIOTIC MEMBRANEQ2

EPITHELIAL CELLSQ2

IMMUNOPHENOTYPEQ2

Abstract

Background: Human amniotic EPITHELIAL CELLS (hAECs) have been introduced as a valuable source of STEM CELLs with therapeutic applications. Despite the extensive study and emerging evidence, cell surface marker signature of hAECs remained controversial. The aim of the present study was to establish an efficient and optimized isolation procedure of hAECs and their characterization in terms of proliferation and STEM CELL markers.Materials and methods: Four human PLACENTAs were collected from healthy women undergoing elective caesarian delivery at term labor under sterile condition. AMNIOTIC MEMBRANEs were carefully isolated and subjected to trypsin digestion. Single cell suspensions were obtained and the epithelial origin was confirmed by assessment of cytokeratin expression. Expression of CD9, CD10, CD29, CD34, CD38, CD44, CD45, CD73, CD105, CD133, MHC-I, HLA-DR, HLA-G, SSEA-4, STRO-1 and OCT-4 was then evaluated by flow cytometry. Results: The overall yield of hAEC ranged from 80-130×106 cells per PLACENTA with more than 98% viability. Microscopic examination revealed that hAECs are large and refractive cells with great capacity to adhere to plastic surfaces. The cells substantially expressed cytokeratin implying their epithelial origin. Flow cytometry data revealed that hAECs are a heterogenic population consisting of immune phenotypically mixed cells. Nearly all purified cells expressed mesenchymal markers, CD9, CD10, CD29, and CD73. A large proportion of cells also expressed HLA-I, HLA-G, STRO-1, OCT-4 and SSEA-4. Isolated cells, however, failed to express CD34, CD38, CD44, CD45, CD105, CD133 and HLA-DR.Conclusion: The procedure presented here is a simple and cost-effective protocol for isolation of a large numbers of viable hAECs. Surface markers and such features as proliferation capacity of hAECs are among the characteristics that are strongly dependent upon isolation and cell culture conditions. Thus such variables should be taken in to consideration when STEM CELL properties of hAECs are to be interpreted.

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APA: Copy

TABATABAEI, MERAJ, ZARNANI, AMIR HOSSEIN, NIKOO, SHOHREH, KESHAVARZI, BAHAREH, RAMZANI TEHRANI, FAHIMEH, AREFI, SOHEILA, MIRZADEGAN, EBRAHIM, & MOSAFFA, NARIMAN. (2013). ISOLATION AND IMMUNOPHENOTYPJNG OF HUMAN AMNIOTIC EPITHELIAL CELLS. PAJOUHESH DAR PEZESHKI, 37(1), 1-7. SID. https://sid.ir/paper/42048/en

Vancouver: Copy

TABATABAEI MERAJ, ZARNANI AMIR HOSSEIN, NIKOO SHOHREH, KESHAVARZI BAHAREH, RAMZANI TEHRANI FAHIMEH, AREFI SOHEILA, MIRZADEGAN EBRAHIM, MOSAFFA NARIMAN. ISOLATION AND IMMUNOPHENOTYPJNG OF HUMAN AMNIOTIC EPITHELIAL CELLS. PAJOUHESH DAR PEZESHKI[Internet]. 2013;37(1):1-7. Available from: https://sid.ir/paper/42048/en

IEEE: Copy

MERAJ TABATABAEI, AMIR HOSSEIN ZARNANI, SHOHREH NIKOO, BAHAREH KESHAVARZI, FAHIMEH RAMZANI TEHRANI, SOHEILA AREFI, EBRAHIM MIRZADEGAN, and NARIMAN MOSAFFA, “ISOLATION AND IMMUNOPHENOTYPJNG OF HUMAN AMNIOTIC EPITHELIAL CELLS,” PAJOUHESH DAR PEZESHKI, vol. 37, no. 1, pp. 1–7, 2013, [Online]. Available: https://sid.ir/paper/42048/en

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