ISOLATION OF MOUSE EBRYONIC STEM CELLS USING VERO CELLS- AS A FEEDER LAYER- AND THEIR IN VITRO DIFFERENTIATION INTO N EURON- LIKE CELLS

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ESMAEILI F.; MOVAHEDIN M.; MOULA SEYED JAVAD; TIRAIHI T.

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Journal: Pathobiology Research Year:2004 | Volume:7 | Issue:1 Page(s): 1-12

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Title

ISOLATION OF MOUSE EBRYONIC STEM CELLS USING VERO CELLS- AS A FEEDER LAYER- AND THEIR IN VITRO DIFFERENTIATION INTO N EURON- LIKE CELLS

Author(s)

ESMAEILI F. | MOVAHEDIN M. | MOULA SEYED JAVAD | TIRAIHI T. | Issue Writer Certificate

Keywords

EMBRYONIC STEM CELLSQ4

VERO CELL LINEQ4

FEEDER CELL LAYERQ4

RETINOIC ACIDQ4

NEURAL DIFFERENTIATIONQ3

Abstract

Purppose: Embryonic stem (ES) cells are pluripotent cells conventionally isolated from the inner call mass of preimplantation embryos. ES cells can be maintained as stable, proliferative and undifferentiated cell lines if cultured in the presence of leukemia inhibitory factor (LIP) or on feeder layers (primary embryonic fibroblasts or STO fibroblasts). Feeder layers synthesize and secrete LIP into the culture medium. Since, based on several evidences, LIP is secreted by mitomycin pretreated Vero cells (a cell line isolated from the green monkey kidney epithelial cells), the main goal of this study was to demonstrate that ES cells can be isolated by using Vero cells as a feeder layer. The criteria used for the evaluation of undifferentiated ES cell phenotype in this study were morphology, evaluation of alkaline phosphatase activity, formation of embryoid bodies (Esb) and NEURAL DIFFERENTIATIONMaterial and Methods: Fully expanded blastocysts were obtained by mating superovulated NMRI female mice and cultured individually on mitomycin C- inactivated Vero cells. After 4-5 days in the culture, The ICMs were plucked off and partially disaggregated in a 0.25% trypsin/EDT A solution. Three to four days later the compact ES cells colonies could be identified. The colonies were dissociated and reseeded on a freshly-treated Vero feeder layer. The cultures were scanned periodically and all the ES cell colonies were passsaged sequentially in the three days intervals. Alkaline phosphatase was detected histochemically, following the fixation of the colonies with 100% ethanol- using α- naphtyl phosphate as a substrate. The undifferentiated mouse ES cells derived on the Vero cells in our laboratory were induced to differentiation using RETINOIC ACID by the 4-/4+ protocol. Several days later, some of the resulting cells posses a neuron-like cell morphology.Results: The resulting cells had high ratio of nucleus to cytoplasm, prominent nucleuli, and a colony morphology similar to that of the murine ES cells. The ES cell colonies expressed the cell surface markers that characterize the undifferentiated ES cells, including alkaline phosphatase.Conclusion: We concluded that Vero cells as a feeder layer may present an alternative avenue in obtaining pluripotent ES cell lines. Although, the application of Verocells in the ES cell culture still requires further investigation.

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APA: Copy

ESMAEILI, F., MOVAHEDIN, M., MOULA, SEYED JAVAD, & TIRAIHI, T.. (2004). ISOLATION OF MOUSE EBRYONIC STEM CELLS USING VERO CELLS- AS A FEEDER LAYER- AND THEIR IN VITRO DIFFERENTIATION INTO N EURON- LIKE CELLS. PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES), 7(1), 1-12. SID. https://sid.ir/paper/81233/en

Vancouver: Copy

ESMAEILI F., MOVAHEDIN M., MOULA SEYED JAVAD, TIRAIHI T.. ISOLATION OF MOUSE EBRYONIC STEM CELLS USING VERO CELLS- AS A FEEDER LAYER- AND THEIR IN VITRO DIFFERENTIATION INTO N EURON- LIKE CELLS. PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES)[Internet]. 2004;7(1):1-12. Available from: https://sid.ir/paper/81233/en

IEEE: Copy

F. ESMAEILI, M. MOVAHEDIN, SEYED JAVAD MOULA, and T. TIRAIHI, “ISOLATION OF MOUSE EBRYONIC STEM CELLS USING VERO CELLS- AS A FEEDER LAYER- AND THEIR IN VITRO DIFFERENTIATION INTO N EURON- LIKE CELLS,” PATHOBIOLOGY RESEARCH (MODARES JOURNAL OF MEDICAL SCIENCES), vol. 7, no. 1, pp. 1–12, 2004, [Online]. Available: https://sid.ir/paper/81233/en

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