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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 E8 medium on vitronectin). Recombinant vitronectin is a functionally defined substrate that supports hPSCs self-renewal through αVβ5 integrin. αVβ1 has been described as a receptor for vitronectin through Ras/MEK/MAPK signaling, and kinases are also stimulated by integrin ligation. This culture system does not need albumin; as collagen I and HSPG do not concentrate on FGF2, it is necessary to have a high concentration of FGF2 in the culture medium.
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f4: Signaling pathways in serum-free and feeder cell-free culture systems (using E8 medium on vitronectin). Recombinant vitronectin is a functionally defined substrate that supports hPSCs self-renewal through αVβ5 integrin. αVβ1 has been described as a receptor for vitronectin through Ras/MEK/MAPK signaling, and kinases are also stimulated by integrin ligation. This culture system does not need albumin; as collagen I and HSPG do not concentrate on FGF2, it is necessary to have a high concentration of FGF2 in the culture medium.

Mentions: Recombinant vitronectin is a functionally defined substrate that supports hPSCs self-renewal through αVβ5 integrin36 (Fig. 4). αVβ5 activates latent TGF-β.114 Chen et al. enhanced the attachment and survival of hPSCs on vitronectin-coated surfaces by excising the N-terminal and/or C-terminal fragments of full-length vitronectin.32 It has also been reported that vitronectin and one of its variants (VTN-NC) support the maintenance of hPSCs through αVβ5 integrin.32,36,115


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 E8 medium on vitronectin). Recombinant vitronectin is a functionally defined substrate that supports hPSCs self-renewal through αVβ5 integrin. αVβ1 has been described as a receptor for vitronectin through Ras/MEK/MAPK signaling, and kinases are also stimulated by integrin ligation. This culture system does not need albumin; as collagen I and HSPG do not concentrate on FGF2, it is necessary to have a high concentration of FGF2 in the culture medium.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: Signaling pathways in serum-free and feeder cell-free culture systems (using E8 medium on vitronectin). Recombinant vitronectin is a functionally defined substrate that supports hPSCs self-renewal through αVβ5 integrin. αVβ1 has been described as a receptor for vitronectin through Ras/MEK/MAPK signaling, and kinases are also stimulated by integrin ligation. This culture system does not need albumin; as collagen I and HSPG do not concentrate on FGF2, it is necessary to have a high concentration of FGF2 in the culture medium.
Mentions: Recombinant vitronectin is a functionally defined substrate that supports hPSCs self-renewal through αVβ5 integrin36 (Fig. 4). αVβ5 activates latent TGF-β.114 Chen et al. enhanced the attachment and survival of hPSCs on vitronectin-coated surfaces by excising the N-terminal and/or C-terminal fragments of full-length vitronectin.32 It has also been reported that vitronectin and one of its variants (VTN-NC) support the maintenance of hPSCs through αVβ5 integrin.32,36,115

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