Optimization of culture conditions in HS5 stroma suppported hematopoietic culture.

• Main Author: Goh, Carlye Qian Ling.
• Other Authors: Mayasari Lim
• Format: Final Year Project
• Language: English
• Published: 2013
• Subjects: DRNTU::Engineering::Bioengineering
• Online Access: http://hdl.handle.net/10356/54647
• Institution: Nanyang Technological University

Stroma cells play an essential role in the bone marrow niche, the environment where hematopoietic cells and progenitors self-renew and differentiate into multiple lineage. A simple 2D model comprising of a monolayer of stroma cells is being used to produce platelets. This study aims to optimize the cell culture by studying HS5 stromal and exogenous growth factor TPO influence on megakaryocytopoiesis. In the first part, growth profile was conducted for HS5 stroma. In the second part of this project, we studied co-cultures consisting of HS5 stroma and/or TPO. The cell population, viability, morphology and phenotype of loose cells were characterized. Under the microscope, significant morphology changes were seen with TPO but the quantitative expression of both megakaryocyte CD 41α and CD42β indicated that culture with both stroma monolayer and TPO has a higher megakaryocyte expression. Culture with stroma monolayer or/and TPO also point to a reduced CD13 and GPA erythroid expression.