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Isolation of embryonic-like stem cell from human placental villi

Yu-Ching Lin1, Chiung-Hsin Chang2, Chen-Hsiang Yu2 and Lynn L. Huang1, 3, 4, 5
  • 1 National Cheng Kung University, Institute of Biotechnology, Taiwan
  • 2 National Cheng Kung University, Department of Obstetrics and Gynecology, Taiwan
  • 3 National Cheng Kung University, Institute of Clinical Medicine, Taiwan
  • 4 National Cheng Kung University, Advanced Optoelectronic Technology Center, Taiwan
  • 5 National Cheng Kung University, Research Center of Excellence in Regenerative Medicine, Taiwan

Introduction: Stem cells have characteristics of differential potential, and embryonic stem cells (ESCs) could be differentiated into three kinds of layers, including ectoderm, mesoderm, and endoderm, hence defined pluripotent[1]. However, the source of embryonic stem cell involved some ethical problem[2]. Therefore, the placental villi belonging to earlier stage of development may still contain some cells which are undifferentiated. The aim of this study was to prove that the cells isolated from placental villi are similar to embryonic stem cells and we referred to these cells as embryonic-like stem cells (ESC-like). Furthermore, to avoid tumorigenesis, mesenchymal stem cells (MSCs) from the same placenta were isolated as feeder layer.

Materials and Methods: 

Isolation of embryonic-like stem cells and mesenchymal stem cells. Amniotic membrane and decidua membrane of placenta were removed, and chorionic villi were cut into pieces. Tissue were digested in collagenase solution at 37℃ water bath with shaking. Digested tissue mixture was filtered through mesh, cell mixture seperated through Percoll gradient. Finally, mononuclear cells were obtained.

Colony formation of embryonic-like stem cells. Cells isolated from placental villi were seed on TCS surface and mesenchymal stem cells used as feeder layer. After one to two weeks, colony formed on the two kinds of surface.

Immunofluorescence staining. Cell were fixed by 4% paraformaldehyde in PBS, blocked using 1% BSA. Cells were incubated with antibody overnight at 4℃, and counterstained with DAPI.

Results and Discussion: In our study, embryonic-like stem cells are capable of colony formation on TCS surface (Fig 1). But these colony differentiated during passage. Therefore, mesenchymal stem cells from the same placenta were utilized as feeder layer to keep embryonic-like stem cells undifferentiated. Secondly, embryonic-like stem cells exhibited the cell markers of embryonic stem cell, including SSEA-3, SSEA-4, TRA-1-60, TRA-1-81 and NANOG (Fig 2). Thirdly, embryonic-like stem cells have the ability to differentiate into three kinds of layers. Based on these result, embryonic-like stem cells which isolated from placental villi were proven to be similar to embryonic stem cells. The source of placental villi is a kind of medical waste which can solve the ethical problem. Embryonic-like stem cells are powerful for utilization in the field of tissue engineering. Based on these pluripotent characteristics, embryonic-like stem cells can be combined with suitable biomaterials and applied to various other research fields.

Fig. 1. Embryonic-like stem cells are capable of colony formation on TCS surface. But these colony differentiated during passage. (B) is the magnified pictograph of (A).

Fig. 2. Embryonic-like stem cells exhibited the cell markers of embryonic stem cell, including (A) TRA-1-81, (B) TRA-1-60 and (C) SSEA-4.

Conclusion: It was found  that the cells which referred to embryonic-like stem cells have pluripotent potential and can be applied to various research fields, in particularly tissue engineering.

The work was supported by grants NSC 99-3111-B-006-002 and MOST 104-2622-B-006-014 from the Ministry of Science and Technology

References:
[1] Alterations of proliferative and differentiation potentials of human embryonic stem cells during long-term culture.
[2] The role of nanotechnology in induced pluripotent and embryonic stem cells research.

Keywords: Cell Differentiation, Tissue Engineering

Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.

Presentation Type: Poster

Topic: Biomaterials in mesenchymal and hematopoietic stem cell biology

Citation: Lin Y, Chang C, Yu C and Huang LL (2016). Isolation of embryonic-like stem cell from human placental villi. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02717

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Received: 27 Mar 2016; Published Online: 30 Mar 2016.