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Related Experiment Video

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Isolation of Human Mesenchymal Stem Cells and their Cultivation on the Porous Bone Matrix
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Processing methods for human amniotic membrane as scaffold for tissue engineering with mesenchymal stromal human

L Echarte1, G Grazioli2, L Pereira3

  • 1Área Terapia Celular y Medicina Regenerativa (ATCMR), Departamento Básico de Medicina, Hospital de Clínicas, Facultad de Medicina, UdelaR, Montevideo, Uruguay.

Cell and Tissue Banking
|July 29, 2022
PubMed
Summary

This study compared processed human amniotic membranes (hAM) as scaffolds for human stromal cells. Decellularized amniotic membrane (DEAM10) showed higher cell proliferation rates compared to frozen amniotic membrane (FEAM).

Keywords:
Amniotic membraneBrain dead donorHuman dental pulp stem cellHuman mesenchymal stromal cellScaffoldTissue engineering construct

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Area of Science:

  • Tissue engineering
  • Biomaterials science
  • Cell biology

Background:

  • Human amniotic membrane (hAM) is a promising biomaterial for tissue regeneration.
  • Different processing methods affect hAM properties and suitability for cell seeding.
  • Investigating novel cell sources like mesenchymal stromal cells (MSCs) is crucial for developing effective tissue constructs.

Purpose of the Study:

  • To compare chemically and physically processed hAM scaffolds.
  • To analyze the cytocompatibility and proliferation rate (PR) of human Dental Pulp Stem Cells (hDPSC) and human Mesenchymal Stromal Cells (hMSC) on hAM scaffolds.
  • To evaluate novel hMSC from bone marrow as a cell source for tissue engineering constructs.

Main Methods:

  • Human amniotic membranes were processed into frozen (FEAM), denuded (DEAM20/10), and decellularized (DAM) forms.
  • Scaffolds were characterized using electron microscopy and X-ray diffraction, with FEAM and DEAM10 selected for cell seeding.
  • Primary hDPSC and hMSC were seeded onto FEAM and DEAM10 scaffolds, and cytotoxicity and PR were assessed in vitro.

Main Results:

  • Four in vitro constructs were developed with no signs of cytotoxicity.
  • Both hDPSC and hMSC demonstrated growth on FEAM and DEAM10 scaffolds.
  • The DEAM10 scaffold facilitated a significantly higher proliferation rate (PR) for both cell types compared to FEAM.

Conclusions:

  • Chemically processed hAM, specifically DEAM10, provides a superior scaffold for supporting human stromal cell growth and proliferation compared to FEAM.
  • The study validates the use of hDPSC and a novel source of hMSC from bone marrow for tissue engineering applications using hAM scaffolds.
  • DEAM10 scaffolds hold potential for developing advanced tissue-engineered constructs with enhanced cellular responses.