1. The document describes a study that isolated hepatic stem cells from human fetal liver tissue and characterized them by examining expression of liver-specific markers. 2. CD34-positive stem cells were isolated from fetal liver cells using magnetic sorting. These cells were characterized for viability, proliferation rate, and expression of alpha-fetoprotein, albumin, and cytokeratins 18 and 19. 3. The results demonstrate that the isolated CD34-positive cells expressed markers characteristic of hepatic stem cells, showing their potential for use in research and cell-based therapies for liver diseases.

1. Isolation of Human hepatic stem cells and characterization by using α - feto protein, Cytokeratin-19 & Albumin Presented by S. Ezhil Mangai INTERNAL GUIDE : L.G. MOHANA DIVYA Lecturer, Department of biotechnology, KSR college of Technology. EXTERNAL GUIDE : DR. ALEEM KHAN, Scientist, Department of stem cell therapy, CLRD.

16. Results and Discussion: 1. Fetal Dissection Ventral view of human fetal liver before perfusion. 1 , 2 , 3 , 4 – Liver lobes. 5 – Hepatic and portal vein. 6- Hepatic lymph nodes. 7- gall bladder.

17. 2. MACS Magnetic assorted cell sorter Pre-running column with saline Collection of CD34-ve cells Column is separated from magnetic field and Eluted with PBS. CD34+ve cells separated.

18. 3 . Viability Test ( Trypan Blue Staining ). Total number of cells = 2368cells/50 μ l Viable cells = 1600cells/50 μ l Dead cells = 768cells/50 μ l Total number of cells in area = 23.68X10 9 %viability = 67.56% Trypan blue staining of CD34+ve cells from human fetal liver cells.

19. 4 . Cell Counting Cells Cell concentration Before MACS After MACS Total cells Positive cells Negative cells 5 million/ml 1 million/ml 4 million/ml

20. 5. Immunocytochemistry FITC conjugated CD34+ve cells under Fluorescent microscope CD34+ve cells

21. 6 . MTT Assay Percentage of activity ( μ g/ml) OD at 580nm 35 39 0.1349 0.1790

22. 7. Gene Expression Analysis (AFP , ALB , CK -18 , CK -19)