Hydrogels are frequently utilized as three-dimensional matrices for the culture and regeneration of soft tissues, but one challenge with the existing hydrogels is that, though the natural extracellular matrix of tissues may be ordered, there are few biocompatible ways to incorporate anisotropy within hydrogels. Liquid crystalline (LC) polymers are well suited for this because of their combination of molecular ordering and polymer elasticity; however, the hydrophobic nature of LC monomers has hindered their polymerization into hydrogels under cytocompatible conditions. This work reports on the generation of main-chain LC hydrogels in aqueous media and the ability of LC phases to affect mesenchymal stem cell behavior. The synthesis results in high gel fraction materials, and calorimetry, thermomechanical analysis, and X-ray scattering show that the networks organize into LC phases in the dry and hydrogel states. Human mesenchymal stem cells (hMSCs) cultured within the hydrogels show excellent viability, and hMSC proliferation proceeds at a faster rate in LC hydrogels compared to non-LC hydrogels. TThe result is a new synthetic approach for calamitic liquid crystalline hydrogels, which support the encapsulation and culture of human stem cells and are expected to enable applications as anisotropic and responsive substrates for tissue engineering and regenerative medicine.