111 International Journal of Animal Biotechnology, Vol.1, No.1 (Dec. 2011) Full length research article Studies of various microbial contaminants during culturing of amniotic fluid derived cells in buffalo (Bubalus bubalis) Kapil Dev a , Shiv Kumar Giri a , Anil Kumar a , Anil Khuttan a , Anita Yadav a , R.P. Mandhan a , Manoj Kumar b , Dheeraj Mohania c , Sushil Kumar Phuilia d , Laxman Singh e , Vijay Kumar f , Sanjeev Kumar a Department of Biotechnology, Kurukshetra University Kurukshetra, Haryana, India b Department of Microbiology and Immunology National Institute of Nutrition, Ministry of Health and Human welfare, Indian council of Medical Research, Hyderabad, India c Sir Ganga Ram Hospital, New Delhi, India d Division of Buffalo Physiology and Reproduction, Central Institute for Research on Buffaloes, Hisar, India e Department of Animal Husbandry and Dairy, RBF College, Bichpuri, Agra, India f Equine Production Campus, Bikaner, India Accepted 5 Oct.2011, Available online 1 Dec. 2011 Abstract Cell culture complexity is as shared as that of culture failure due to contamination. All cell culture in laboratories and cell culture workers have acknowledged it. Culture contaminants may be organic or elementary, seen or hidden, unhelpful or apparently gentle, but in all gear they harmfully influence both the use of cell cultures and the excellence of research work. Major microbial contaminants in the cell culture include Staphylococcus epidermidis, S. aureus, Bacillus subtilis, Escherichia. coli, Mycoplasma orale, M. fermentans and Saccharomyces cerevisiae. There are several factors which influence the amniotic fluid (AF) cell culture like level of blood cells, volume of amniotic fluid, attachment time, number and microbial contamination. Amniotic fluid derived cells in buffalo (Bubalus bubalis) were cultured in normal suitable media and effects of microbial contaminants were studied. Keywords: Amniotic fluid cell, Microorganism, Buffalo, Cell culture 1. Introduction 1 The majority noticeable result of cell culture contamination is the failure of time, money and attempt tired rising cultures and location up experiments. However, the fewer obvious cost are often more serious. First there are unfavorable special effects on cultures suffering from hidden chemical or organic (biological) contaminants. These unseen contaminants can attain elevated densities changing the development and individuality of the cultures. Worse, yet are the potentially incorrect or mistaken results obtained by innocently working with these cryptically impure cultures. Harvest, such as vaccines, drugs or monoclonal antibodies, manufactured by these cultures will possibly be ineffective. For a number of researchers the harshest outcome of contamination is suffering the discomfiture and damage to their status that results when they inform collaborators or journals that their investigational results are defective and have to be retracted due to contaminants in their cultures. Preventing all cell culture contamination * Corresponding author: sgautam@kuk.ac.in has long been the dream of many researchers, but it is an impractical, if not impossible, dream. It has been reported that AF could be a promising source of putative stem cells, and that the AF cells are easily reprogrammable than other somatic cells (Li et al., 2009; Galende et al., 2010). Ethical problems and material sourcing, which limit embryonic stem cell research lead to isolation of cells from human amniotic fluid and have been named them human amniotic fluid stem (AFS) cells (Ryan ,1998) thereby providing a new route for stem cell research (Chiavegato et al., 2007). The information on AFS cells in buffalo species is very rare and has been reported from a few labs only (Yadav et al., 2011; Dev et al., 2012a, b). In the present investigation, effect of various microbial contaminants that influence the primary AF cell culture have been studied. 2. Materials and methods Chemicals and media