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Human adipose-derived stem cell isolation from fat tissues by membrane filtration method via nylon net filters having different pore sizes

Hong-Ren Lin1, Saradaprasan Muduli1, Yi Tung Lu1 and Akon Higuchi1, 2
  • 1 National Center University, Chemical engineering and material science, Taiwan
  • 2 College of Science, King Saud University, Department of Botany and Microbiology, Saudi Arabia

Human adult stem cells, such as human adipose-derived stem cells (hADSCs), are considered to be a more attractive source of stem cells than human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). This is because human adult stem cells do not generate the ethical concerns that accompany hESCs. The hADSCs exhibit heterogeneous characteristics, indicating various genotypes and differentiation abilities. The isolated hADSCs can possess different purity levels and divergent properties depending on the purification methods used[1]. In order to make sure the importance and difference between different pore sizes, we decided the nylon mesh filter with pore size from 11 to 60 mm.

Fig. 1 hADSCs purification by membrane migration method.

We developed a hybrid-membrane migration method that purifies hADSCs from a fat tissue solution with extremely high purity and pluripotency. Digest fat tissue with 0.75% collagenese for 1 hour, obtained a primary fat-tissue solution and permeated it through the porous membranes with a pore size from 11 to 60 mm by the membrane filtration method, and the membranes were incubated in cell culture medium for 15-18 days to expand cell number and run flow-cytometer and qRT-PCT to analysis purity and pluripotent gene expression.

Fig. 2 Purity of hADSCs after filtration and purified by membrane migration method through 11, 20, 41 and 60 µm pore size of nylon mesh filter.

The hADSCs that migrated from the membranes contained an extremely high percentage (e.g., >90%) of cells positive for mesenchymal stem cell markers and the cells isolated by membrane filtration method showed  higher expression of some pluripotency genes (Oct4, Sox2, klf4 and Nanog) compared with the cells isolated using the conventional culture method. When pore size is smaller then 20 mm, it could remain high purity hADSCs. The smaller pore size had higher purity and pluripotency.

References:
[1] A. Higuchi and C.-T. Wang, et al., A hybrid-membrane migration method to isolate high-purity adipose-derived stem cells from fat tissues, Sci Rep., 5 (2015) 10217.

Keywords: biomaterial, microstructure, material design

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

Presentation Type: Poster

Topic: Biomaterials for mechanical interfaces

Citation: Lin H, Muduli S, Lu Y and Higuchi A (2016). Human adipose-derived stem cell isolation from fat tissues by membrane filtration method via nylon net filters having different pore sizes. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.01755

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