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Nanotopographical control of human embryonic stem cell differentiation into definitive endoderm
Author(s) -
Ghanian Mohammad Hossein,
Farzaneh Zahra,
Barzin Jalal,
Zandi Mojgan,
KazemiAshtiani Mohammad,
Alikhani Mehdi,
Ehsani Morteza,
Baharvand Hossein
Publication year - 2015
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.35483
Subject(s) - endoderm , embryonic stem cell , microbiology and biotechnology , extracellular matrix , regenerative medicine , materials science , cellular differentiation , stem cell , tissue engineering , biology , gene , genetics
Derivation of definitive endoderm (DE) from human embryonic stem cells (hESCs) can address the needs of regenerative medicine for endoderm‐derived organs such as the pancreas and liver. Fibrous substrates which topographically recapitulate native extracellular matrix have been known to promote the stem cell differentiation. However, the optimal fiber diameter remains to be determined for the desired differentiation. Here, we have developed a simple method to precisely fabricate electrospun poly(ε‐caprolactone) fibers with four distinct average diameters at nano‐ and microscale levels (200, 500, 800, and 1300 nm). Human ESCs were cultured as clumps or single cells and induced into DE differentiation to determine the optimal topography leading to the promoted differentiation compared with planar culture plates. Gene expression analysis of the DE‐induced cells showed significant upregulation of DE‐specific genes exclusively on the 200‐nm fibers. By Western blot analysis, significant expression of DE‐specific proteins was found when hESCs were cultured on the 200 nm substrate as single cells rather than clumps, probably due to more efficient cell–matrix interaction realized by morphological observations of the cell colonies. The results indicated that nanofibrillar substrates, only at ultrathin fiber diameters, provided a better environment for DE differentiation of hESC, which holds great promise in prospective tissue engineering applications. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 3539–3553, 2015.