title: Efficient generation of hepatic cells from mesenchymal stromal cells by an innovative bio-microfluidic cell culture device.
abstract: Mesenchymal stromal cells (MSCs) are multipotent and have great potential in cell therapy. Previously we reported the differentiation potential of human MSCs into hepatocytes in vitro and that these cells can rescue fulminant hepatic failure. However, the conventional static culture method neither maintains growth factors at an optimal level constantly nor removes cellular waste efficiently. In addition, not only is the duration of differentiating hepatocyte lineage cells from MSCs required to improve, but also the need for a large number of hepatocytes for cell therapy has not to date been addressed fully. The purpose of this study is to design and develop an innovative microfluidic device to overcome these shortcomings.
  
  
  
  We designed and fabricated a microfluidic device and a culture system for hepatic differentiation of MSCs using our protocol reported previously. The microfluidic device contains a large culture chamber with a stable uniform flow to allow homogeneous distribution and expansion as well as efficient induction of hepatic differentiation for MSCs.
  
  
  
  The device enables real-time observation under light microscopy and exhibits a better differentiation efficiency for MSCs compared with conventional static culture. MSCs grown in the microfluidic device showed a higher level of hepatocyte marker gene expression under hepatic induction. Functional analysis of hepatic differentiation demonstrated significantly higher urea production in the microfluidic device after 21 days of hepatic differentiation.
  
  
  
  The microfluidic device allows the generation of a large number of MSCs and induces hepatic differentiation of MSCs efficiently. The device can be adapted for scale-up production of hepatic cells from MSCs for cellular therapy.
authors.affiliation: Institute of Clinical Medicine, National Yang-Ming University, No. 155, Sec. 2, Linong Street, Taipei, 112, Taiwan (Republic of China).
authors.forename: Meng-Hua
authors.initials: MH
authors.lastname: Yen
authors.affiliation: Institute of Clinical Medicine, National Yang-Ming University, No. 155, Sec. 2, Linong Street, Taipei, 112, Taiwan (Republic of China).
authors.forename: Yuan-Yi
authors.initials: YY
authors.lastname: Wu
authors.affiliation: Stem Cell Research Center, National Yang-Ming University, No. 155, Sec. 2, Linong Street, Taipei, 112, Taiwan (Republic of China).
authors.forename: Yi-Shiuan
authors.initials: YS
authors.lastname: Liu
authors.affiliation: Taiwan International Graduate Program, National Yang Ming University and Academia Sinica, No. 155, Sec. 2, Linong Street, Taipei, 112, Taiwan (Republic of China).
authors.forename: Marilyn
authors.initials: M
authors.lastname: Rimando
authors.affiliation: Center for Stem Cell Research, Wan Fang Hospital, Taipei Medical University, No. 111, Section 3, Hsing-Long Rd, Taipei, 116, Taiwan (Republic of China). wh9801@yahoo.com.tw.|Graduate Institute of Clinical Medicine, Taipei Medical University, No. 250 Wuxing Street, Taipei City, 110, Taiwan (Republic of China). wh9801@yahoo.com.tw.|Department of Ophthalmology, Wan Fang Hospital, Taipei Medical University, No. 111, Sec. 3, Hsing-Long Rd, Taipei, 116, Taiwan (Republic of China). wh9801@yahoo.com.tw.
authors.forename: Jennifer Hui-Chun
authors.initials: JH
authors.lastname: Ho
authors.affiliation: Institute of Clinical Medicine, National Yang-Ming University, No. 155, Sec. 2, Linong Street, Taipei, 112, Taiwan (Republic of China). DAV47@tpech.gov.tw.|Department of Medical Research, Taipei Veterans General Hospital, No. 201, Sec. 2, Shipai Rd, Beitou District, Taipei City, 112, Taiwan (Republic of China). DAV47@tpech.gov.tw.|Taipei City Hospital, No. 145, Zhengzhou Rd, Datong Dist., Taipei, 103, Taiwan (Republic of China). DAV47@tpech.gov.tw.
authors.forename: Oscar Kuang-Sheng
authors.initials: OK
authors.lastname: Lee
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D019070:Cell Lineage / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D015870:Gene Expression / Q000502:physiology; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008810:Mice, Inbred C57BL; D044085:Microfluidics / Q000379:methods*
chemical_list: D015415:Biomarkers
keywords: Hepatic differentiation; Hepatocyte; Mesenchymal stromal cells; Microfluidic device