title: Locust bean gum as an alternative polymeric coating for embryonic stem cell culture.
abstract: Pluripotent embryonic stem cells (ESCs) have self-renewal capacity and the potential to differentiate into any cellular type depending on specific cues (pluripotency) and, therefore, have become a vibrant research area in the biomedical field. ESCs are usually cultured in gelatin or on top of a monolayer of feeder cells such as mitotically inactivated mouse embryonic fibroblasts (MEFsi). The latter is the gold standard support to maintain the ESCs in the pluripotent state. Examples of versatile, non-animal derived and inexpensive materials that are able to support pluripotent ESCs are limited. Therefore, our aim was to find a biomaterial able to support ESC growth in a pluripotent state avoiding laborious and time consuming parallel culture of MEFsi and as simple to handle as gelatin. Many of the new biomaterials used to develop stem cell microenvironments are using natural polymers adsorbed or covalently attached to the surface to improve the biocompatibility of synthetic polymers. Locust beam gum (LBG) is a natural, edible polymer, which has a wide range of potential applications in different fields, such as food and pharmaceutical industry, due to its biocompatibility, adhesiveness and thickening properties. The present work brings a natural system based on the use of LBG as a coating for ESC culture. Undifferentiated mouse ESCs were cultured on commercially available LBG to evaluate its potential in maintaining pluripotent ESCs. In terms of morphology, ESC colonies in LBG presented the regular dome shape with bright borders, similar to the colonies obtained in co-cultures with MEFsi and characteristic of pluripotent ESC colonies. In short-term cultures, ESC proliferation in LBG coating was similar to ESC cultured in gelatin and the cells maintained their viability. The activity of alkaline phosphatase and Nanog, Sox2 and Oct4 expression of mouse ESCs cultured in LBG were comparable or in some cases higher than in ESCs cultured in gelatin. An in vitro differentiation assay revealed that mouse ESCs cultured in LBG preserve their tri-lineage differentiation capacity. In conclusion, our data indicate that LBG coating promotes mouse ESC growth in an undifferentiated state demonstrating to be a viable, non-animal derived alternative to gelatin to support pluripotent mouse ESCs in culture.
authors.affiliation: Regenerative Medicine Program, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, Portugal; IBB - Institute for Biotechnology and Bioengineering, Centre for Molecular and Structural Biomedicine (CBME), Universidade do Algarve, Portugal; PhD Program in Biomedical Sciences, Universidade do Algarve, Portugal.
authors.forename: Ana Rubina
authors.initials: AR
authors.lastname: Perestrelo
authors.affiliation: IBB - Institute for Biotechnology and Bioengineering, Centre for Molecular and Structural Biomedicine (CBME), Universidade do Algarve, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: Grenha
authors.affiliation: Centro de Investigação em Química do Algarve (CIQA) & Departamento de Química e Farmácia, Faculdade de Ciências e Tecnologia, Universidade do Algarve, Portugal.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Rosa da Costa
authors.affiliation: Regenerative Medicine Program, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, Portugal; IBB - Institute for Biotechnology and Bioengineering, Centre for Molecular and Structural Biomedicine (CBME), Universidade do Algarve, Portugal; Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Campo Mártires da Pátria 130, 1169-056 Lisboa, Portugal. Electronic address: jose.belo@fcm.unl.pt.
authors.forename: José António
authors.initials: JA
authors.lastname: Belo
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D019070:Cell Lineage; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology; D005685:Galactans / Q000737:chemistry / Q000494:pharmacology*; D018398:Homeodomain Proteins / Q000378:metabolism; D008351:Mannans / Q000737:chemistry / Q000494:pharmacology*; D051379:Mice; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D053149:Plant Gums / Q000737:chemistry / Q000494:pharmacology*; D011108:Polymers / Q000737:chemistry; D055748:SOXB1 Transcription Factors / Q000378:metabolism
chemical_list: D005685:Galactans; D018398:Homeodomain Proteins; D008351:Mannans; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D050814:Octamer Transcription Factor-3; D053149:Plant Gums; D011108:Polymers; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D000469:Alkaline Phosphatase; C017471:locust bean gum
keywords: Embryonic stem cell culture; Gelatin substitute growth support; Locust bean gum; Natural polymers; Pluripotency