The umbilical cord tissue has gained attention in recent years as a source of multipotent cells. Due to its widespread availability, the umbilical cord may be an excellent alternative source of cells for regenerative medicine. Anatomically, umbilical cord tissue is constituted of several different parts, and, accordingly, immunostaining of cord tissue sections revealed differential distribution of several markers and extracellular matrix, distinguishing the various layers. Wharton's jelly is the major component filling the inner part of the umbilical cord tissue, and it has been commonly used as a source of obtaining multipotent cells from umbilical cord. We recently reported isolating mesenchymal stem cells from cord lining membrane (sub-amnion). However, because of several anatomically distinct zones found in the umbilical cord, isolated multipotent cells sometimes show heterogeneity. In addition, differences in isolation technique may lead to further variation. In this review, we discuss the similarities and differences between the cells derived from each sub-region, including sub-amnion as recently reported by us. We further explore the specific features and advantages/disadvantages of Wharton's jelly and the other sub-compartments in the umbilical cord tissue as sources of stem cells/multipotent cells.