CHAPTER I INTRODUCTION The use of amniotic fluid to assess the ...

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2 already be near to senescence. This research effort isolates cells by cloning for homogeneity to retrieves the more unique and stable continuous amniotic cell lines. When a new cell line has derived, it is necessary to evaluate details of its origin and expressed features. This aspect of cell culture is a crucial important with the widespread dissemination of a cell line through research laboratories and cell banks. In particular if a cell line becomes incorporated into a research that requires validation of its components, then the authentications of the cell line get necessary. It is, therefore, vital that detailing the origin and handling of the cell lines are kept from the time of isolation and periodically during use. The purpose of a cell line to be characterized are depending on the situation of use such as; to confirm the species of origin, to correlate with the tissue origin, to determine the cell line transformation, to demonstrate the cross-contamination, to indicate the prone of genetic instability and phenotypic variation, and to identify a specific cell line with in a group from the same origin (Butler and Dowson, 1992; Freshney, 2000). Several detailed protocols to success of the above purpose are widely available (Kuchler, 1977; Masters, 2000; Freshney, 2000). However, some basic characteristics are studied in this research for the newly form cell lines and their sub-clones including; morphology (type of cells in cultures), plating efficiency (the percentage of cells seeded at subculture that gives rise to colony), seeding efficiency (the percentage of the inoculums that attaches to the substrate within a stated period of time), growth pattern or growth curve (a semilogarithmic plot of the cell number on a logarithmic scale against time on a linear scale for a proliferating cell culture), viability (survival of the cells) and the karyotyping (the distinctive chromosomal complement of cells). These characteristics will be very useful for any future research with the continuous cell lines. Amniotic cells containing many of cell types present in the original tissue. A wide variety of investigations performed since the 1980s have provided evidences that cells of all three germ layers (ectoderm, mesoderm and endoderm) can be detected in human amniotic fluid (Hoenh and Salk, 1982 and Gosden, 1983). Despite, the widespread and well-established use of amniotic cells in routine prenatal genetic testing, current knowledge about the origin and properties of these cells is limited (Prusa and Hengstschläger, 2002). Recently, some reports mentioned that the amniotic fluid contained a variety of stem cells, which were shed from embryonic and
3 extra-embryonic tissues during the process of fetal development and growth (Prusa et al., 2003; Fauza, 2004; Tsai et al., 2004). Amniotic cells could transform into several cells and may be the source of powerful therapy for human congenital disorder and as a potential source of pluripotent stem cells (Kakishita, 2003 and Tsai et al., 2005). In addition, Delo et al. (2006) mentioned about number of promising novel therapeutic concepts using these cells in tissue engineering, cell transplantation and gene therapy. Current stem cell based approaches could be employed in cellreplacement therapy or drug treatments. Thus, amniotic cells are intriguing as a possible source of pluripotent stem cells for cell-based therapeutics, and do not raise the ethical concerns associated with use of embryonic stem cells (Kim et al., 2007). Furthermore, many experiments carried out in vitro are for the sole purpose of determining the potential cytotoxicity of the compounds being studied. Either because the compounds are being used as pharmaceuticals or cosmetics or must be shown to be nontoxic or designed as anticancer agents and cytotoxicity may be crucial to their action (Freshney, 2000). In this study, the two new amniotic cell lines are studied for their potential use in cytotoxic assay. The response of these cells to a standard component of pharmaceutical and cosmetic compound will be provided with the effective inhibitory concentrations. OBJECTIVES This study was initiated with the aims to; (i) Establish the homogenous amniotic cell lines from amniotic fluid of Thais by cell cloning. (ii) Study some characteristics and examine the Stemness of the developed cell lines. (iii) Study the toxicity response potential of the cell lines in cytotoxicity assay.
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CHAPTER I INTRODUCTION The use of amniotic fluid to assess the ...

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