105 Pakistan Veterinary Journal ISSN: 0253-8318 (PRINT), 2074-7764 (ONLINE) Accessible at: www.pvj.com.pk Effect of Somatic Cell Types and Culture Medium on in vitro Maturation, Fertilization and Early Development Capability of Buffalo Oocytes H. Jamil*, H. A. Samad, N. Rehman, Z. I. Qureshi and L. A. Lodhi Department of Theriogenology, University of Agriculture, Faisalabad, Pakistan *Corresponding author: drhjamil@hotmail.com ARTICLE HISTORY ABSTRACT Received: Revised: Accepted: July 12, 2010 September 30, 2010 October 14, 2010 Key words: Buffalo oocytes In vitro maturation In vitro fertilization Somatic cells co culture This study was designed to evaluate the efficacy of different somatic cell types and media in supporting in vitro maturation (IVM), in vitro fertilization (IVF) and early embryonic development competence of buffalo follicular oocytes. Cumulus oocyte complexes were collected for maturation from follicles (>6mm) of buffalo ovaries collected at the local abattoir. Oocytes were co-cultured in tissue culture medium (TCM-199) with either granulosa cells, cumulus cells, or buffalo oviductal epithelial cells (BOEC) @ 3x10 6 cells/ml or in TCM-199 without helper cells (control) at 39°C and 5%CO 2 in humidified air. Fresh semen was prepared in modified Ca ++ free Tyrode medium. Fertilization was carried out in four types of media: i) Tyrode lactate albumin pyruvate (TALP), ii) TALP+BOEC, iii) modified Ca ++ free Tyrode and iv) modified Ca ++ free Tyrode+BOEC. Fertilized oocytes were cultured for early embryonic development in TCM-199 with and without BOEC. Higher maturation rates were observed in the granulosa (84.24%) and cumulus cells (83.44%) than BOEC co culture system (73.37%). Highest fertilization rate was obtained in modified Ca ++ free Tyrode with BOEC co culture (70.42%), followed by modified Ca ++ free Tyrode alone (63.77%), TALP with BOEC (36.92%) and TALP alone (10.94%). Development of early embryos (8-cell stage) improved in TCM- 199 with BOEC co culture than TCM-199 alone. From the results of this study, it can be concluded that addition of somatic cells (granulosa cells, cumulus cells) results in higher maturation rates of buffalo follicular oocytes than BOEC co culture system, while fertilization rate improved in modified Ca ++ free Tyrode with and without BOEC. Addition of BOEC to TCM-199 improved the developmental capacity of early embryo. ©2011 PVJ. All rights reserved To Cite This Article: Jamil H, HA Samad, N Rehman, ZI Qureshi and LA Lodhi, 2011. Effect of somatic cell types and culture medium on in vitro maturation, fertilization and early development capability of buffalo oocytes. Pak Vet J, 31(2): 105-108. INTRODUCTION Buffalo is the main dairy animal in Pakistan and accounts for approximately 75% of all milk produced in the country. Inherent reproductive problems, such as delayed puberty (Nandi et al., 2002), delayed first calving (Lundstrom et al., 1982), late postpartum conception (Arora and Jain, 1988) and longer calving intervals (Singh and Roy, 1996) lead to low reproduction and production performance. Genetic improvement in this important animal resource is focused on its role as a major provider of milk and meat in this part of the world. Limitation to genetic improvement imposed by inherent biological parameters can be overcome by the use of recently developed reproductive biotechnologies. In vitro production of buffalo embryos adopting the procedures developed for cattle has received increasing interest in recent years. The efficiency of in vitro embryo production (IVEP) in buffalo is much lower than in cattle in terms of IVM, IVF and yield of transferable quality embryos (Madan et al., 1994; Palta and Chauhan, 1998). Nandi et al. (2002) reported that the in vitro embryo production system in buffalo is suboptimal and requires substantial improvement along with several other problems that need to be resolved before in vitro embryo production technology can be used regularly in buffalo breeding. In mammals, somatic cell-oocytes communication provides beneficial effects by providing nutritional molecules that are necessary for growth and development and by transmitting signals that regulate oocytes maturation RESEARCH ARTICLE