We present an optimization method to retrieve the gradient index (GRIN) distribution of the in-vitro crystalline lens from optical path difference data extracted from OCT images. Three-dimensional OCT images of the crystalline lens are obtained in two orientations (with the anterior surface up and posterior surface up), allowing to obtain the lens geometry. The GRIN reconstruction method is based on a genetic algorithm that searches for the parameters of a 4-variable GRIN model that best fits the distorted posterior surface of the lens. Computer simulations showed that, for noise of 5 μm in the surface elevations, the GRIN is recovered with an accuracy of 0.003 and 0.010 in the refractive indices of the nucleus and surface of the lens, respectively. The method was applied to retrieve three-dimensionally the GRIN of a porcine crystalline lens in vitro. We found a refractive index ranging from 1.362 in the surface to 1.443 in the nucleus of the lens, an axial exponential decay of the GRIN profile of 2.62 and a meridional exponential decay ranging from 3.56 to 5.18. The effect of GRIN on the aberrations of the lens also studied. The estimated spherical aberration of the measured porcine lens was 2.87 μm assuming a homogenous equivalent refractive index, and the presence of GRIN shifted the spherical aberration toward negative values (-0.97 μm), for a 6-mm pupil.