title: Purification of human adipose-derived stem cells from fat tissues using PLGA/silk screen hybrid membranes.
abstract: The purification of human adipose-derived stem cells (hADSCs) from human adipose tissue cells (stromal vascular fraction) was investigated using membrane filtration through poly(lactide-co-glycolic acid)/silk screen hybrid membranes. Membrane filtration methods are attractive in regenerative medicine because they reduce the time required to purify hADSCs (i.e., less than 30 min) compared with conventional culture methods, which require 5-12 days. hADSCs expressing the mesenchymal stem cell markers CD44, CD73, and CD90 were concentrated in the permeation solution from the hybrid membranes. Expression of the surface markers CD44, CD73, and CD99 on the cells in the permeation solution from the hybrid membranes, which were obtained using 18 mL of feed solution containing 50 × 10⁴ cells, was statistically significantly higher than that of the primary adipose tissue cells, indicating that the hADSCs can be purified in the permeation solution by the membrane filtration method. Cells expressing the stem cell-associated marker CD34 could be successfully isolated in the permeation solution, whereas CD34⁺ cells could not be purified by the conventional culture method. The hADSCs in the permeation solution demonstrated a superior capacity for osteogenic differentiation based on their alkali phosphatase activity, their osterix gene expression, and the results of mineralization analysis by Alizarin Red S and von Kossa staining compared with the cells from the suspension of human adipose tissue. These results suggest that the hADSCs capable of osteogenic differentiation preferentially permeate through the hybrid membranes.
authors.affiliation: Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan 32001, Taiwan; Taiwan Landseed Hospital, 77, Kuangtai Road, Pingjen City, Taoyuan 32405, Taiwan.
authors.forename: Da-Chung
authors.initials: DC
authors.lastname: Chen
authors.affiliation: Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan 32001, Taiwan.
authors.forename: Li-Yu
authors.initials: LY
authors.lastname: Chen
authors.affiliation: Institute of Systems Biology and Bioinformatics, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan 32001, Taiwan; Cathay Medical Research Institute, Cathay General Hospital, No. 32, Ln 160, Jian-Cheng Road, Hsi-Chi City, Taipei 221, Taiwan.
authors.forename: Qing-Dong
authors.initials: QD
authors.lastname: Ling
authors.affiliation: Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan 32001, Taiwan.
authors.forename: Meng-Hsueh
authors.initials: MH
authors.lastname: Wu
authors.affiliation: Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan 32001, Taiwan.
authors.forename: Ching-Tang
authors.initials: CT
authors.lastname: Wang
authors.affiliation: Department of Medical Microbiology and Parasitology, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia.
authors.forename: S
authors.initials: S
authors.lastname: Suresh Kumar
authors.affiliation: Department of Chemical Engineering, R&D Center for Membrane Technology, Chung Yuan Christian University, 200, Chung-Bei Rd., Chungli, Taoyuan 320, Taiwan.
authors.forename: Yung
authors.initials: Y
authors.lastname: Chang
authors.affiliation: Department of Botany and Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia.
authors.forename: Murugan A
authors.initials: MA
authors.lastname: Munusamy
authors.affiliation: Department of Botany and Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia.
authors.forename: Abdullah A
authors.initials: AA
authors.lastname: Alarfajj
authors.affiliation: Hungchi Women & Children's Hospital, No. 233, Yuanhua Rd., Jhongli, Taoyuan 320, Taiwan.
authors.forename: Han-Chow
authors.initials: HC
authors.lastname: Wang
authors.affiliation: Taiwan Landseed Hospital, 77, Kuangtai Road, Pingjen City, Taoyuan 32405, Taiwan.
authors.forename: Shih-Tien
authors.initials: ST
authors.lastname: Hsu
authors.affiliation: Department of Chemical and Materials Engineering, National Central University, No. 300, Jhongda RD., Jhongli, Taoyuan 32001, Taiwan; Cathay Medical Research Institute, Cathay General Hospital, No. 32, Ln 160, Jian-Cheng Road, Hsi-Chi City, Taipei 221, Taiwan; Department of Botany and Microbiology, College of Science, King Saud University, Riyadh 11451, Saudi Arabia; Department of Reproduction, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo 157-8535, Japan. Electronic address: higuchi@ncu.edu.tw.
authors.forename: Akon
authors.initials: A
authors.lastname: Higuchi
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002452:Cell Count; D002454:Cell Differentiation / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D002469:Cell Separation; D005374:Filtration; D005434:Flow Cytometry; D006801:Humans; D019344:Lactic Acid / Q000494:pharmacology*; D008567:Membranes, Artificial; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D008875:Middle Aged; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects / Q000235:genetics; D011100:Polyglycolic Acid / Q000494:pharmacology*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D047011:Silk / Q000494:pharmacology*; D012996:Solutions; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D017154:Stromal Cells / Q000187:drug effects / Q000378:metabolism; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism
chemical_list: D015415:Biomarkers; D008567:Membranes, Artificial; D047011:Silk; D012996:Solutions; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid
keywords: Adipose-derived stem cells; Bioseparation; Differentiation; Membrane purification; Osteoblast; Poly(lactide-co-glycolic acid)