Serum-Free Defined Media

The growth medium for cells grown in culture is routinely supplemented with animal and/or human serum to optimize and enhance cell viability. The constituents of serum include water, amino acids, glucose, albumins, immunoglobulins, and one or more bioactive agents. Potential bioactive agents present in serum include agents that induce proliferation, agents that accelerate phenotypic expression, agents that induce differentiation, agents that inhibit proliferation, agents that inhibit phenotypic expression, and agents that inhibit differentiation. Unfortunately, the identity(ies), concentration(s), and potential combinations of specific bioactive agents contained in different lots of serum is/are unknown. One or more of these unknown agents in serum have shown a negative impact on the isolation, cultivation, cryopreservation, and purification of lineage-uncommitted blastomere-like stem cells. To circumvent the problems inherent with the use of serum, the Company's scientists have developed proprietary serum-free defined (SFD) reagents for the isolation, cultivation, cryopreservation, and purification of lineage-uncommitted blastomere-like stem cells.

With addition of specific bioactive factors to the Company's base media, the BLSCs grown in a specific medium are induced to differentiate to a specific germ-layer lineage that will eventually give rise to progenitor cells of a specific tissue phenotype. For example, it has been reported in the scientific literature that growth factors such as EGF (epidermal growth factor) and FGF-2 (fibroblast growth factor-2) are needed in the tissue culture medium for propagating -islet progenitor cells of the pancreas. Similarly, by adding unique components to the Company's SFD medium, a proprietary -islet pancreatic culture medium kit can be used to generate insulin-producing and glucose-responsive cells in vitro with the BLSCs. The end-user will have an invaluable research tool to interrogate these cells for novel pathways in their research and drug discovery programs for treating diabetes.

The Company will also develop proprietary SFD medium for growing embryonic stem cells. With minor modification to its BLSC propagation medium, the Company expects to develop SFD for both human and mouse embryonic stem cells. Researchers are currently propagating human embryonic stem cells using culture conditions requiring serum and, for human embryonic stem cells, a mouse monolayer which apparently secretes autocrine and paracrine factors that inhibits differentiation of human embryonic stem cells in culture. In order to propagate undifferentiated mouse embryonic stem cells, the culture medium requires the presence of a leukemia inhibitory factor (LIF) that apparently activates the STAT3 molecular signaling pathway that maintains their pluripotency or "stemness."