Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by the gradual loss of motor neurons, leading to paralysis and ultimately death. Multiple pathways contribute to neuronal death and play a role in both the development and progression of the disease. This complexity makes effective treatment particularly challenging. Adipose-derived stem cell-conditioned medium (ASC-CM) is a biologically derived reagent rich in neuroprotective and neurotrophic factors that has been shown to exhibit neuroprotective effects in both animal and cell culture models of neurodegenerative diseases. It is essential for the survival, growth, and maintenance of specific types of neurons and could serve as an ideal treatment for neurodegenerative diseases, which are often associated with hormone or growth factor deficiencies. Our previous studies evaluated the therapeutic effects of ASC-CM on SOD1G93A mice, a widely studied animal model of ALS. In this chapter, we provide guidance on adipose-derived stem cell preparation, conditioned medium collection, administration of ASC-CM in SOD1 mice, and methods to monitor and evaluate its effectiveness.