Ascorbic acid enhances the cardiac differentiation of induced pluripotent stem cells through promoting the proliferation of cardiac progenitor cells

Abstract

Generation of induced pluripotent stem cells (iPSCs) has opened new avenues for the investigation of heart diseases, drug screening and potential autologous cardiac regeneration. However, their application is hampered by inefficient cardiac differentiation, high interline variability, and poor maturation of iPSC-derived cardiomyocytes (iPS-CMs). To identify efficient inducers for cardiac differentiation and maturation of iPSCs and elucidate the mechanisms, we systematically screened sixteen cardiomyocyte inducers on various murine (m) iPSCs and found that only ascorbic acid (AA) consistently and robustly enhanced the cardiac differentiation of eleven lines including eight without spontaneous cardiogenic potential. We then optimized the treatment conditions and demonstrated that differentiation day 2-6, a period for the specification of cardiac progenitor cells (CPCs), was a critical time for AA to take effect. This was further confirmed by the fact that AA increased the expression of cardiovascular but not mesodermal markers. Noteworthily, AA treatment led to approximately 7.3-fold (miPSCs) and 30.2-fold (human iPSCs) augment in the yield of iPS-CMs. Such effect was attributed to a specific increase in the proliferation of CPCs via the MEK-ERK1/2 pathway by promoting collagen synthesis. In addition, AA-induced cardiomyocytes showed better sarcomeric organization and enhanced responses of action potentials and calcium transients to β-adrenergic and muscarinic stimulations. These findings demonstrate that AA is a suitable cardiomyocyte inducer for iPSCs to improve cardiac differentiation and maturation simply, universally, and efficiently. These findings also highlight the importance of stimulating CPC proliferation by manipulating extracellular microenvironment in guiding cardiac differentiation of the pluripotent stem cells.

Vitamin C induces periodontal ligament progenitor cell differentiation via activation of ERK pathway mediated by PELP1

“…ERK activity is recognized as the decision maker during the process of embryonic stem cells differentiation (Kunath et al, 2007). And by activating ERK, VC enhances the cardiac differentiation of iPSCs and induces the apoptosis of acute myeloid leukemia cells (Park et al, 2005;Cao et al, 2012). So, whether ERK was involved in VCinduced osteogenic differentiation of PDL progenitor cells became our concern.…”

“…It is an indispensable supplement for promoting the proliferation and differentiation of a variety of mesenchymal stem cells by enhancing collagen biosynthesis, ALP activity, the expression of BSP and other extracellular matrix production (Franceschi et al, 1994;Shiga et al, 2003;Wang et al, 2006;Cao et al, 2012;Mekala et al, 2013). Evidences also indicate that VC can induce iPSCs (induced pluripotent stem cells) differentia-& tion and malignant cells apoptosis via ERK/MAPK pathway (Park et al, 2005;Cao et al, 2012). ERK/MAPK pathway lies in the central position in mesenchymal stem cells differentiation and skeletal development (Lai et al, 2001;Ge et al, 2007).…”

The 2013 special issue on stem cell biology

“…Along the way, the field of stem cell biology has naturally become an important research area covered by papers published in Cell Research [2]. For instance, in the past year of 2012, Cell Research has published a number of important papers related to the stem cell field, covering diverse aspects and topics such as induced pluripotent stem (iPS) cells [3][4][5][6][7], mechanistic studies of pluripotency and differentiation [8][9][10][11], modeling of human diseases using stem cell-based systems [6,12], direct reprogramming of somatic cells to other cell types without passing through an pluripotent intermediate [13][14][15][16], as well as neural crest stem cells [17] and cancer stem cells [18][19][20].…”

We heart cultured hearts. A comparative review of methodologies for targeted differentiation and maintenance of cardiomyocytes derived from pluripotent and multipotent stem cells

“…Supplementation with ascorbic acid greatly increases the yield of cardiomyocytes from human iPSCs (> 30-fold) [163]. However, the authors of the latter study report that there was a 'window' during which the application of ascorbic acid was effective (within the timeframe of specification of cardiac progenitors).…”