Hepatic differentiation from human mesenchymal stem cells on a novel nanofiber scaffold

Abstract

Abstract:The emerging fields of tissue engineering and biomaterials have begun to provide potential treatment options for liver failure. The goal of the present study is to investigate the ability of a poly L-lactic acid (PLLA) nanofiber scaffold to support and enhance hepatic differentiation of human bone marrow-derived mesenchymal stem cells (hMSCs). A scaffold composed of poly L-lactic acid and collagen was fabricated by the electrospinning technique. After characterizing isolated hMSCs, they were seeded onto PLLA nanofiber scaffolds and induced to differentiate into a hepatocyte lineage. The mRNA levels and protein expression of several important hepatic genes were determined using RT-PCR, immunocytochemistry and ELISA. Flow cytometry revealed that the isolated bone marrow-derived stem cells were positive for hMSC-specific markers CD73, CD44, CD105 and CD166 and negative for hematopoietic markers CD34 and CD45. The differentiation of these stem cells into adipocytes and osteoblasts demonstrated their multipotency. Scanning electron microscopy showed adherence of cells in the nanofiber scaffold during differentiation towards hepatocytes. Our results showed that expression levels of liver-specific markers such as albumin, α-fetoprotein, and cytokeratins 8 and 18 were higher in differentiated cells on the nanofibers than when cultured on plates. Importantly, liver functioning serum proteins, albumin and α-1 antitrypsin were secreted into the culture medium at higher levels by the differentiated cells on the nanofibers than on the plates, demonstrating that our nanofibrous scaffolds promoted and enhanced hepatic differentiation under our culture conditions. Our results show that the engineered PLLA nanofibrous scaffold is a conducive matrix for the differentiation of MSCs into functional hepatocyte-like cells. This represents the first step for the use of this nanofibrous scaffold for culture and differentiation of stem cells that may be employed for tissue engineering and cell-based therapy applications.

The effects of poly L-lactic acid nanofiber scaffold on mouse spermatogonial stem cell culture

“…PLLA nanofibers composed of PLLA and collagen fabricated by the electrospinning technique were purchased from Stem Cells Technology (Tehran, Iran). 31 The PLLA nanofiber was used in a culture system with both cryopreserved and fresh SSCs. After placing the nanofibers on the dishes, fresh and frozen-thawed spermatogonial cells were seeded (5 × 10 5 cells) on nanofiber and cultured in three groups: (1) fresh cells, (2) frozen-thawed cells, and (3) cells obtained from frozenthawed testis tissue.…”

“…[23][24][25][26] Poly L-lactic acid (PLLA) is one of the most promising biodegradable, biocompatible, and US Food and Drug Administration-approved polymers 27 and can easily be electrospun to form a 3D non-woven network. 28,29 To date, the proliferation of muscle-derived stem cells 30 and the differentiation of hepatic cells from human mesenchymal stem cells 31 have been established on PLLA. 3D soft agar culture system and electrospun polyamide nanofiber (Ultra-Web™; Corning Life Sciences, Tewksbury, MA, USA) also have been reported to yield complete in vitro spermatogenesis of mouse testicular germ cells 32,33 as well as short-term culture of spermatogonial stem-like cell colonies.…”

A synthetic nanofibrillar matrix promotes in vitro hepatic differentiation of embryonic stem cells and induced pluripotent stem cells

“…In addition, the ECM was found to deposit as an extensive scaffold on the basal surface of the cells attached to NFs (Shih et al, 2006;Chua et al, 2007;Ma et al, 2008). Importantly, the sNF system has been reported to enhance not only the differentiation of murine ES cells into neural lineages (Lim et al, 2010;Purcell et al, 2012;Xie et al, 2009), but also differentiation from human MSCs into hepatoblasts (Ghaedi et al, 2012;Kazemnejad et al, 2009).…”

New Models for the In Vitro Study of Liver Toxicity: 3D Culture Systems and the Role of Bioreactors

“…Examples of the sophisticated models that, for various purposes, have been obtained from primary hepatocytes, are those generated by using bio-degradable nano-structured substrates (Kim et al, 1998), heat-sensitive polymers (Ohashi et al, 2007), various 3D matrices (Fiegel et al, 2008;ZQ. Feng et al, 2010;Ghaedi et al, 2012), or synthetic self-assembling hydrogels (Wang et al, 2008).…”