This chapter aims to present a brief discussion on bioengineered human endometrium in vitro. The entire procedure has to be performed in a class 10,000 tissue culture laboratory, well equipped with air management by air handling unit (AHU). The cells handling should be performed in a laminar flow with class 100 atmosphere. All the disposable ware should be made up of tissue culture grade polypropelene of BD falcon make and the culture media used should be from Sigma-Aldrich. The procedure is contraindicated in patients with genital tuberculosis, Asherman’s syndrome where the amount of endometrium obtained for culture is usually inadequate. Human endometrium is a layered tissue with basically three components, which are defined in this chapter. As such, the procedure of culture does not have any complications. However, sometimes the contamination in the culture system can prevent the cell proliferation and on some occasions may spoil the entire culture with resultant degeneration of all the cells.