Human skin has a complex multilayer structure consisting of non-homogeneous, non-linear, viscoelastic and anisotropic materials subjected to in vivo natural pre-tension. This natural tension stems from networks of collagen and elastin fibers. The 3D organization of the collagen and elastin fibers underpins the multidirectional natural tensions in the skin volume while the state of the networks formed influences the surface topography of the skin. This topography depends on the area of the body and on the age of the person. Experiments reported in the literature have been performed ex vivo or on cadavers. By contrast, this work proposes the characterization of the anisotropic natural tension of the human skin in vivo. Experimental tests were performed on the forearms and thighs of 42 female volunteers representing two age groups [20 - 30] and [45-55] years old. Non-contact impact tests and skin-folding tests were conducted using devices developed at the LTDS (Lyon, France). The impact test generated a Rayleigh wave that spread in the skin. The speed of this wave was measured in 7 directions to study the anisotropy of the skin tension. The image of the skin relief at rest and during the skin folding test was reconstructed by optical confocal microscopy and provided the density of the skin lines printed on the outer surface of the skin. Skin folding test enables the clinician's manual procedure to be instrumented to identify tension lines i.e., Langer lines, for better healing during a surgical procedure. The main directions of natural skin tension deduced from the measured wave speed and the densities of skin lines were [40°-60°] for the forearm and [0°-20°] for the thigh, considering that the longitudinal axis of the body is situated at 90° and the transversal axis at 0°. This method shows the remarkable effect of age and body area on the mechanical behavior of human skin in vivo. The elastic properties and natural tension of the skin decrease with age. This decrease is greater in the directions orthogonal to the skin's tension lines, leading to the accentuation of the anisotropic behavior of the cutaneous tissue. The main direction of skin tension is highly dependent on the area of the body and is directed towards a preferred direction which corresponds to the main direction of skin tension.