This dataset contains Raman spectra, each acquired from an individual, living, cell entrapped within a soft or stiff gelatin methacrylate hydrogel or from a cell-free region of the hydrogel sample. Spectra were acquired from the following cell types: Madin-Darby Canine Kidney cell (MDCK); Chinese hamster ovary cell (CHO-K1); transfected CHO-K1 cell that expressed the SNAP-tag and HaloTag reporter proteins fused to an organelle-specific protein (CHO-T); human monocyte-like cell (THP-1); inactive macrophage-like (M0-like); active anti-inflammatory macrophage-like (M2-like), pro/anti-inflammatory macrophage-like (M1/M2-like). These spectra are useful for identifying whether the hydrogel matrix obscures the Raman spectral signatures that are characteristic of each of these cell types.