Ab212166

Enzyme-linked immunosorbent assay (ELISA; ab212166, Abcam, Cambridge, UK) was used according to the manufacturer’s instructions to assess the exosomal BDNF level. Each sample was assessed in duplicate, and the concentration of BDNF was compared with the standard sample provided.

Acetylcholinesterase (AChE) activities and BDNF contents were measured in hippocampus using commercial colorimetric AChE activity assay kit (#ab138871, Abcam, CA, and #ab212166, Abcam) according to the manufacturer's protocol.

Subdomains of executive function including the task-switching ability and inhibiting interference were assessed by the Trail Making Test (TMT) [29 (link)] and Stroop Color and Word Test (SCWT) [30 (link)], respectively. The difference in times to complete TMT part B and part A (TMT B-A) was used to reflect the switching ability between tasks [29 (link)]. For SCWT, the number of correct answers in the word (W), color (C), and color-word (CW) conditions were used to determine the interference score (IG) following the formula: IG = CW − [(W × C)/(W + C)] [30 (link)]. A lower IG score indicates a greater difficulty in inhibiting interference. In addition, episodic memory was assessed using a logical memory test [31 (link)]. The scores of delayed recall from two short stories were collected. A higher score indicates a greater performance of memory. For plasma brain-derived nerve growth factor (BDNF) levels, a commercially available quantitative sandwich enzyme-linked immunosorbent assay (ELISA) kit was used for BDNF measurement, following standard protocols provided by the test manufacturer (ab212166, Abcam).

The major GFs and NFs related to the retinal cell protection and regeneration in AMMSC-NCM and AMMSC-ERCM were determined via ELISA, according to the manufacturer’s instructions. Briefly, AMMSC-NCM or AMMSC-ERCM were put into the ELISA wells with antibodies specific for FGF (ab219636; Abcam), EGF (ab217772; Abcam), IGF (ab211652; Abcam), VEGF (ab222510; Abcam), TGF-β (ab100647; Abcam), BDNF (ab212166; Abcam), or PDGF (ab100622; Abcam) and incubated at room temperature for 0.5–1 h. After washing 3–4 times with wash buffer, the primary antibodies were added, and reacted at room temperature for 1 h and then washed again. Secondary antibodies were treated and incubated at room temperature for 30–45 min, washed, and substrates were applied to develop color for 10–30 min. After stopping the color development with a stop solution, the absorbance was measured at 450 nm.