Counteracting apoptosis and necrosis with hypoxia responsive expression of Bcl-2Delta.

In the encapsulated environment of biohybrid artificial organs, cells often encounter a deficiency in oxygen, which lead to apoptosis, necrosis, and lost of productivity. Two vectors with constitutive CMV promoters were constructed to examine the ability of Bcl-2Delta to help C2C12 mouse myoblasts maintain exogenous protein production under hypoxia. Two additional vectors with hypoxia-inducible promoters (5HRE) that switched on Bcl-2Delta expression based on low oxygen levels (0.0%, 0.5%, 1.0%, 2.0%, 5.0%, or 21.0%) were tested for protein productivity and protection against hypoxic stresses. A yellow fluorescent protein was used as a model protein in all vector constructs. C2C12 cells with Bcl-2Delta consistently produced more protein regardless of the oxygen level or promoter used. Cells utilizing the 5HRE rather than the CMV promoter showed an increased level of protein production as the oxygen was decreased. Among the cells with 5HRE promoters, the presence of Bcl-2Delta also increased viability and decreased apoptosis.