Noggin maintains pluripotency of human embryonic stem cells grown on Matrigel

Abstract

Objective Spontaneous differentiation of human embryonic stem cell (hESC) cultures is a major concern in stem cell research. Physical removal of differentiated areas in a stem cell colony is the current approach used to keep the cultures in a pluripotent state for a prolonged period of time. All hESCs available for research require unidentified soluble factors secreted from feeder layers to maintain the undifferentiated state and pluripotency. Under experimental conditions, stem cells are grown on various matrices, the most commonly used being Matrigel. Materials and Methods We propose an alternative method to prevent spontaneous differentiation of hESCs grown on Matrigel that uses low amounts of recombinant noggin. We make use of the porosity of Matrigel to serve as a matrix that traps noggin and gradually releases it into the culture to antagonize bone morphogenetic proteins (BMP). BMPs are known to initiate differentiation of hESCs and are either present in the conditioned medium or are secreted by hESCs themselves. Results hESCs grown on Matrigel supplemented with noggin in conditioned medium from feeder layers (irradiated mouse embryonic fibroblasts) retained both normal karyotype and markers of hESC pluripotency for 14 days. In addition, these cultures were found to have increased cell proliferation of stem cells as compared to hESCs grown on Matrigel alone. Conclusion Noggin can be utilized for short term prevention of spontaneous differentiation of stem cells grown on Matrigel.

RNA-Binding Protein FXR2 Regulates Adult Hippocampal Neurogenesis by Reducing Noggin Expression

“…Noggin plays important roles in many types of stem cells and helps maintain pluripotency in cultured stem cells (Chambers et al, 2009; Chaturvedi et al, 2009). With regard to adult neurogenesis, Noggin inhibits BMP signaling to promote NPC proliferation and neuronal differentiation, while inhibiting glial differentiation (Chmielnicki et al, 2004; Lim et al, 2000).…”

Exogenous Activation of BMP‐2 Signaling Overcomes TGFβ‐Mediated Inhibition of Osteogenesis in Marfan Embryonic Stem Cells and Marfan Patient‐Specific Induced Pluripotent Stem Cells

“…Thus, in both of these cases, BMP signaling appears to suppress Wnt signaling in the context of stem cell niches. Several reports also indicated that noggin, an inhibitor of BMP signaling, maintains pluripotency of human stem cells [55–57]. Conversely, it has been demonstrated that TGFβ1 induces rapid nuclear translocation of β‐catenin in adult MSCs in a Smad3‐dependent manner leading to their proliferation [54].…”

Scaffold Materials for hES Cell Culture and Differentiation

“…11 The highly complex and heterogeneous nature of Matrigel 12 means that its exact composition varies from batch to batch, making it less than ideal for hESC culture. 16 However, each of these systems utilized Matrigel as a substrate, and there are considerable efforts underway to replace Matrigel with a defined substrate. 13 To generate hESC lines for clinical applications, the cells should ideally be cultured in fully defined conditions.…”