The Significance Application of Indigenous Phytohemagglutinin (PHA) Mitogen on Metaphase and Cell Culture Procedure

Abstract

Phytohemagglutinin (PHA) is a lectin, obtained from the red kidney bean that binds to the membranes of T-cells and stimulates metabolic activity, cell division, etc. The object of this research was the comparison between self made PHA (Indigenous) and imported commercial one, following conventional and High Resolution Cell Synchronization technique (HRCS) .From each blood sample of healthy individual donor replicate cell culture with two different PHA (self-made and commercial imported) with same concentration were cultured simultaneously. For culture cells, 3-5 × 1066 cells were cultured in 4 mL medium( RPMI 1640 supplemented with 15 per cent heat inactivated fetal bovine serum, 0.1 mL Phytohemagglutinin was added and kept at 37°C in an atmosphere containing 5% CO2. The processing of mitotic division from 48 h and 72 h cultures was performed according to the standard and High Resolution Cell Synchronization technique. Cytogenetic studies were performed in 100 normal healthy blood donor individuals. Statistical analysis was performed by SPSS (version 16, Inc.USA) software.Our results indicate that the preparation of fresh Phytohemagglutinin at the time of cell division and cell culture procedure reveals satisfactory score. The overall frequency of mitotic index in our study was better when compared with commercial imported Phytohemagglutinin (p < 0.001).The significant differences in the results may be due to fresh preparation. However, cost effective, easy and nearest approach of this indigenous product and high demand for this product among health care services can be considered.