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What Kind of Signaling Maintains Pluripotency and Viability in Human-Induced Pluripotent Stem Cells Cultured on Laminin-511 with Serum-Free Medium?

Nakashima Y, Omasa T - Biores Open Access (2016)

Bottom Line: In hiPSCs, the interaction of laminin-511/α6β1 integrin with the cell-cell adhesion molecule E-cadherin confers protection against apoptosis through the Ras homolog gene family member A (RhoA)/Rho kinase (ROCK) signaling pathway (the major pathways for cell death) and the proto-oncogene tyrosine-protein kinase Fyn (Fyn)-RhoA-ROCK signaling pathway.A combination of growth factors, medium constituents, cell membrane-located E-cadherin, and α6β1 integrin-induced signaling is required for pluripotent cell proliferation and for optimal cell survival on a laminin-511 scaffold.In this review, we discuss and explore the influence of growth factors on the cadherin and integrin signaling pathways in serum-free and xeno-free cultures of hiPSCs during the preparation of products for regenerative medicinal therapies.

View Article: PubMed Central - PubMed

Affiliation: Department of Material and Life Science, Graduate School of Engineering, Osaka University , Osaka, Japan .

ABSTRACT
Xeno-free medium contains no animal-derived components, but is composed of minimal growth factors and is serum free; the medium may be supplemented with insulin, transferrin, and selenium (ITS medium). Serum-free and xeno-free culture of human-induced pluripotent stem cells (hiPSCs) uses a variety of components based on ITS medium and Dulbecco's modified Eagle's medium/Ham's nutrient mixture F12 (DMEM/F12) that contain high levels of iron salt and glucose. Culture of hiPSCs also requires scaffolding materials, such as extracellular matrix, collagen, fibronectin, laminin, proteoglycan, and vitronectin. The scaffolding component laminin-511, which is composed of α5, β1, and γ1 chains, binds to α3β1, α6β1, and α6β4 integrins on the cell membrane to induce activation of the PI3K/AKT- and Ras/MAPK-dependent signaling pathways. In hiPSCs, the interaction of laminin-511/α6β1 integrin with the cell-cell adhesion molecule E-cadherin confers protection against apoptosis through the Ras homolog gene family member A (RhoA)/Rho kinase (ROCK) signaling pathway (the major pathways for cell death) and the proto-oncogene tyrosine-protein kinase Fyn (Fyn)-RhoA-ROCK signaling pathway. The expression levels of α6β1 integrin and E-cadherin on cell membranes are controlled through the activation of insulin receptor/insulin, FGF receptor/FGF2, or activin-like kinase 5 (ALK5)-dependent TGF-β signaling. A combination of growth factors, medium constituents, cell membrane-located E-cadherin, and α6β1 integrin-induced signaling is required for pluripotent cell proliferation and for optimal cell survival on a laminin-511 scaffold. In this review, we discuss and explore the influence of growth factors on the cadherin and integrin signaling pathways in serum-free and xeno-free cultures of hiPSCs during the preparation of products for regenerative medicinal therapies. In addition, we suggest the optimum serum-free medium components for use with laminin-511, a new scaffold for hiPSC culture.

No MeSH data available.


Related in: MedlinePlus

Signaling pathways in serum-free and feeder cell-free culture systems (using MEF-CM on Matrigel). Matrigel includes laminin-111, collagen IV, fibronectin, and vitronectin and supports robust hPSC growth. Collagen IV binds to α2β1 integrin and transduces a RhoGAP function to activate PI3K binding to AKT; activation of the latter pathway promotes cell growth in hiPSCs. Vitronectin supports the maintenance of hPSCs through αVβ5 integrin. αVβ1 integrin is able to recognize vitronectin and fibrinogen. Fibronectin can also bind to αVβ1 integrin and α5β1 integrin to promote secretion of laminin-332, -511, -521, and collagen IV from hiPSCs. hPSC, human pluripotent stem cells; MEF-CM, MEF-conditioned medium.
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f2: Signaling pathways in serum-free and feeder cell-free culture systems (using MEF-CM on Matrigel). Matrigel includes laminin-111, collagen IV, fibronectin, and vitronectin and supports robust hPSC growth. Collagen IV binds to α2β1 integrin and transduces a RhoGAP function to activate PI3K binding to AKT; activation of the latter pathway promotes cell growth in hiPSCs. Vitronectin supports the maintenance of hPSCs through αVβ5 integrin. αVβ1 integrin is able to recognize vitronectin and fibrinogen. Fibronectin can also bind to αVβ1 integrin and α5β1 integrin to promote secretion of laminin-332, -511, -521, and collagen IV from hiPSCs. hPSC, human pluripotent stem cells; MEF-CM, MEF-conditioned medium.

Mentions: Activin A has a role in a wide range of cellular processes from cell proliferation and differentiation to apoptosis. Initially, activin A binds to type II activin A receptors (ActRIIA or ActRIIB) and then recruits type IB activin A receptor (ALK4). ALK4 interacts with and phosphorylates the SMAD family members 2 and 3 (SMAD2 and SMAD3). Vallier et al.37–39 demonstrated that the cooperation of the activin A pathway and FGF2 is necessary for the maintenance of pluripotency in hPSCs.40 When active, PI3K/AKT establishes conditions where activin A/SMAD2/3 stimulate self-renewal in hiPSCs by activating target genes, including Nanog.41,42 This stimulatory effect involves the interaction of activin A and SMAD2/3, and may also require upregulation of FGF2 pathways39,43 (Fig. 2).


What Kind of Signaling Maintains Pluripotency and Viability in Human-Induced Pluripotent Stem Cells Cultured on Laminin-511 with Serum-Free Medium?

Nakashima Y, Omasa T - Biores Open Access (2016)

Signaling pathways in serum-free and feeder cell-free culture systems (using MEF-CM on Matrigel). Matrigel includes laminin-111, collagen IV, fibronectin, and vitronectin and supports robust hPSC growth. Collagen IV binds to α2β1 integrin and transduces a RhoGAP function to activate PI3K binding to AKT; activation of the latter pathway promotes cell growth in hiPSCs. Vitronectin supports the maintenance of hPSCs through αVβ5 integrin. αVβ1 integrin is able to recognize vitronectin and fibrinogen. Fibronectin can also bind to αVβ1 integrin and α5β1 integrin to promote secretion of laminin-332, -511, -521, and collagen IV from hiPSCs. hPSC, human pluripotent stem cells; MEF-CM, MEF-conditioned medium.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4834485&req=5

f2: Signaling pathways in serum-free and feeder cell-free culture systems (using MEF-CM on Matrigel). Matrigel includes laminin-111, collagen IV, fibronectin, and vitronectin and supports robust hPSC growth. Collagen IV binds to α2β1 integrin and transduces a RhoGAP function to activate PI3K binding to AKT; activation of the latter pathway promotes cell growth in hiPSCs. Vitronectin supports the maintenance of hPSCs through αVβ5 integrin. αVβ1 integrin is able to recognize vitronectin and fibrinogen. Fibronectin can also bind to αVβ1 integrin and α5β1 integrin to promote secretion of laminin-332, -511, -521, and collagen IV from hiPSCs. hPSC, human pluripotent stem cells; MEF-CM, MEF-conditioned medium.
Mentions: Activin A has a role in a wide range of cellular processes from cell proliferation and differentiation to apoptosis. Initially, activin A binds to type II activin A receptors (ActRIIA or ActRIIB) and then recruits type IB activin A receptor (ALK4). ALK4 interacts with and phosphorylates the SMAD family members 2 and 3 (SMAD2 and SMAD3). Vallier et al.37–39 demonstrated that the cooperation of the activin A pathway and FGF2 is necessary for the maintenance of pluripotency in hPSCs.40 When active, PI3K/AKT establishes conditions where activin A/SMAD2/3 stimulate self-renewal in hiPSCs by activating target genes, including Nanog.41,42 This stimulatory effect involves the interaction of activin A and SMAD2/3, and may also require upregulation of FGF2 pathways39,43 (Fig. 2).

Bottom Line: In hiPSCs, the interaction of laminin-511/α6β1 integrin with the cell-cell adhesion molecule E-cadherin confers protection against apoptosis through the Ras homolog gene family member A (RhoA)/Rho kinase (ROCK) signaling pathway (the major pathways for cell death) and the proto-oncogene tyrosine-protein kinase Fyn (Fyn)-RhoA-ROCK signaling pathway.A combination of growth factors, medium constituents, cell membrane-located E-cadherin, and α6β1 integrin-induced signaling is required for pluripotent cell proliferation and for optimal cell survival on a laminin-511 scaffold.In this review, we discuss and explore the influence of growth factors on the cadherin and integrin signaling pathways in serum-free and xeno-free cultures of hiPSCs during the preparation of products for regenerative medicinal therapies.

View Article: PubMed Central - PubMed

Affiliation: Department of Material and Life Science, Graduate School of Engineering, Osaka University , Osaka, Japan .

ABSTRACT
Xeno-free medium contains no animal-derived components, but is composed of minimal growth factors and is serum free; the medium may be supplemented with insulin, transferrin, and selenium (ITS medium). Serum-free and xeno-free culture of human-induced pluripotent stem cells (hiPSCs) uses a variety of components based on ITS medium and Dulbecco's modified Eagle's medium/Ham's nutrient mixture F12 (DMEM/F12) that contain high levels of iron salt and glucose. Culture of hiPSCs also requires scaffolding materials, such as extracellular matrix, collagen, fibronectin, laminin, proteoglycan, and vitronectin. The scaffolding component laminin-511, which is composed of α5, β1, and γ1 chains, binds to α3β1, α6β1, and α6β4 integrins on the cell membrane to induce activation of the PI3K/AKT- and Ras/MAPK-dependent signaling pathways. In hiPSCs, the interaction of laminin-511/α6β1 integrin with the cell-cell adhesion molecule E-cadherin confers protection against apoptosis through the Ras homolog gene family member A (RhoA)/Rho kinase (ROCK) signaling pathway (the major pathways for cell death) and the proto-oncogene tyrosine-protein kinase Fyn (Fyn)-RhoA-ROCK signaling pathway. The expression levels of α6β1 integrin and E-cadherin on cell membranes are controlled through the activation of insulin receptor/insulin, FGF receptor/FGF2, or activin-like kinase 5 (ALK5)-dependent TGF-β signaling. A combination of growth factors, medium constituents, cell membrane-located E-cadherin, and α6β1 integrin-induced signaling is required for pluripotent cell proliferation and for optimal cell survival on a laminin-511 scaffold. In this review, we discuss and explore the influence of growth factors on the cadherin and integrin signaling pathways in serum-free and xeno-free cultures of hiPSCs during the preparation of products for regenerative medicinal therapies. In addition, we suggest the optimum serum-free medium components for use with laminin-511, a new scaffold for hiPSC culture.

No MeSH data available.


Related in: MedlinePlus