**4. Advantages of organoids over 2D techniques for cancer treatment**

The development of organoid technology has overcome the limitations caused by 2D cell cultures in the following ways: 2D cultures were not repetitive of parent cultures while organoids were a portrait of their parent cells, the lack of predictivity faced in the use of 2D cultures was not visible while using organoid technology, and in 2D cell, culture models waste were being generated from growth medium which was overcome by organoid technology [54]. In organoid technology, multiple cell types were developed *in-vivo* that resemble the parent cells that have enormous applications in cancer research [11]. Tumor modeling is one of the vital applications of organoids as this technology help in identifying toxicity and effect of drugs before administering to patients which is not possible in 2D technology [40]. Organoids have strengthened the understanding of various disease progression and also helped scientists to study the pathways of disease carefully that enhance the treatment methods [55]. Organoids have extended their applications in various fields such as fundamental research [56], disease modeling [57], development of personalized medicine [58], and transplantation [59].

To overcome the technological limitations of cancer treatment, organoids were developed in which ASCs were proliferated in-vitro and self-organized into 3D structures by utilizing extracellular matrix proteins like BME and Matrigel as medium [60]. The main advantage of organoid technology is tissue-specific growth factors can be utilized based on the epithelial cells used for analysis [61]. Based on numerous studies, the ASCs derived organoids were found to be genetically and phenotypically stable and permit the expansion of stem cells in a required path which allows them to differentiate into tissue-specific cells [62]. Apart from tumor heterogeneity, more focus is required on lymphocyte infiltration trafficking along with the clinical response of individual patients to cancer treatment [63].
