Table 1.
Summary of the isolation and validation techniques of milk derived exosomes in literature published since 2017.
| Sample Source | Isolation Strategy | Validation | Comments | Reference |
|---|---|---|---|---|
| Human breast milk | Centrifugation with ExoQuick | Exosomal miR-148a was negatively associated with infant weight, fat mass, and fat free mass, while miR-30b was positively associated with infant weight, percent body fat, and fat mass at 1 month. | Shah et al. (10) | |
| Human breast milk | Ultracentrifugation | NTA, TEM, Western blot (Hsp70, CD9) | Chen et al. (11) | |
| Human breast milk | Centrifugation with exosome isolation kit | TEM, NTA, Western blot (CD81, CD63) | Exosomes prevent necrotizing enterocolitis by reducing inflammation and injury as well as restoring tight junction proteins. | He et al. (12) |
| Cow milk | Ultracentrifugation and size exclusion chromatography | TEM, Western blot (Tsg101, Flot-1, Alix, CD63) | Milk exosomes can be taken up by intestinal epithelial cells and mediated functional intracellular delivery of siRNA. | Warren et al. (13) |
| Cow milk | Ultracentrifugation and size exclusion chromatography | Western blot (Hsp 90, CD63, Tsg101), NTA | Milk exosomes can be sued as nanocarriers of functional miRNAs in RNA-based therapy. | Pozo-Acebo et al. (14) |
| Cow milk | Ultracentrifugation | DLS, SEM, Western blot (CD63, CD9) | Milk exosomes attenuated purine nucleotide catabolism and improved energy status in oxidatively stressed IEC-6 cells | Wang et al. (15) |
| Cow milk | Ultracentrifugation | DLS, TEM, Western blot (CD9, CD63) | Exosomes successfully delivered epicatechin gallate into SHSY5Y cells and exhibited enhanced neuroprotective effects. | Luo et al. (16) |
| Cow milk and colostrum | Ultracentrifugation | Western blot (CD9, CD63milk only, Hsp70) | Exosomes can be taken up by human intestinal epithelia cells, not cytotoxic | Ross et al. (17) |
| Cow milk and yak milk | Ultracentrifugation with rennet precipitation | TEM, DLS, Western blot (CD63, Hsp70, Tsg101) | Milk exosmes alleviated LPS induced intestinal inflammation by inhibiting PI3K/AKT/C3 pathway activation. | Gao et al. (18) |
| Porcine colostrum and mature milk | Ultracentrifugation and SEC | NTA, TEM, Western blot (Tsg101) | The pathways related to homeostasis are upregulated in colostrum exosomes whereas pathways related to endothelia cell development and lipid metabolism are upregulated in milk exosomes. | Ferreira et al. (19) |
| Caprine colostrum and mature milk | Ultracentrifugation | miR-30a-5p, miR-22-3p, and miR-26a are highly conserved in colostrum and mature milk in cows, caprines and humans. | Yun et al. (20) | |
| Colostrum powder | Ultracentrifugation | DLS, NTA, AFM | Exosomes can reduce or completely mitigate the immunotoxicity effects caused by solvent based paclitaxel. | Kandimalla et al. (21) |
| Porcine colostrum and mature milk | Ultracentrifugation and SEC | NTA, TEM, Western blot (Tsg101) | The pathways related to homeostasis are upregulated in colostrum exosomes whereas pathways related to endothelia cell development and lipid metabolism are upregulated in milk exosomes. | Ferreira et al. (19) |
| Cow milk | Ultracentrifugation | DLS, SEM, Western blot (CD63, CD9) | Milk exosomes attenuated purine nucleotide catabolism and improved energy status in oxidatively stressed IEC-6 cells | Wang et al. (15) |
| Human breast milk | Centrifugation with ExoQuick | Exosomal miR-148a was negatively associated with infant weight, fat mass, and fat free mass, while miR-30b was positively associated with infant weight, percent body fat, and fat mass at 1 month. | Shah et al. (10) | |
| Human milk | Ultracentrifugation | NTA, TEM, Western blot (Hsp70, CD9) | Chen et al. (11) | |
| Human breast milk | Centrifugation with exosome isolation kit | TEM, NTA, Western blot (CD81, CD63) | Exosomes prevent necrotizing enterocolitis by reducing inflammation and injury as well as restoring tight junction proteins. | He et al. (12) |
| Cow milk | Ultracentrifugation | DLS, TEM, Western blot (CD9, CD63) | Exosomes successfully delivered epicatechin gallate into SHSY5Y cells and exhibited enhanced neuroprotective effects. | Luo et al. (16) |
| Skim milk | Ultracentrifugation | DLS, TEM, Western blot (CD9, Tsg101, CD63) | Exosomes and miRNA can cross the placenta and promote embryo survival in mice | Sadri et al. (22) |
| Cow milk | Differential centrifugation | DLS, Western blot (Tsg101, CD81, Alix) | Milk exosomes can protect macrophages from chemotherapeutic drug-induced cytotoxicity. | Matic et al. (23) |
| Cow milk | Ultracentrifugation | IL-2 and IL-12 enhanced IFN-γ production with cow milx exosomes but not alone. | Komine-Aizawa et al. (24) | |
| Cow milk | Differential centrifugation | Western blot(CD9, CD63, Tsg101) | Milk exosome based drug delivery system constructed showed controlled drug release, biocompatibility and effective in treating OSCC | Zhang et al. (25) |
| Cow milk | Ultracentrifugation | TEM, NTA, Western blot (Tsg101, Alix, CD9, CD81) | Exosomes can alter murine gut microbiota and SCFA in feces, as well regulate local intestinal immunity. | Tong et al. (26) |
| Canine colostrum | Ultracentrifugation | Western blot (Alix, Tsg101, Hsp70) | Exosomes modified the proliferation and secretory profiles in canine mesenchymal stem cells. | Villatoro et al. (27) |
| Human Breast milk | Centrifugation and filtration with ExoQuick reagent | Electron microscopy, DLS | Human milk-derived exosomes induced proliferation- and epithelial mesenchymal transformation-related changes. MDEs inhibited proliferation and DNMT1 expression in cells with knockdown of miRNA-148a | Reif et al. (28) |
| Cow milk | Ultracentrifugation | DLS, TEM, Western blot (CD63, CD81, ANXA5, FLOT1, ICAM, Tsg101) | Exosomes can be absorbed as intact perticles from the gastrointestinal tract via the Fc receptor. It can be modified with ligands to promote retention in target tissues. | Betker et al. (29) |
| Cow milk | Ultracentrifugation | TEM, DLS, Western blot (CD9, CD63) | Milk exosomes contain lncRNA which participate in immunity, development, and reproduction. LncRNA had different expression patterns during different stages of lactation. | Zeng et al. (30) |
| Cow milk | Ultracentrifugation | NTA, TEM, Flow cytometry (CD63, Hsp70, CD9, CD81) | Milk exosomes prevented experimental NEC-induced intestinal injury by increasing goblet cell production and ER function. | Li et al. (31) |
| Yak milk and cow milk | Ultracentrifugation/ ultracentrifugation with rennet precipitation |
TEM, DLS, Western blot (CD63, Hsp70, Tsg101) | Yak milk exosomes has a more efficient effect on IEC-6 cell growth under hypoxic conditions than cow milk exosomes. | Gao et al. (32) |
| Breast milk | Ultracentrifugation | NTA, Western blot (CD81, clathrin) | Human breast milk-derived exosomes allow IECs to be protected from oxidative stress. | Martin et al. (33) |
| Skim milk | Ultracentrifugation | NTA, TEM, Western blot (CD9, Tsg101, CD63) | Milk exosomes are bioavailable and miRNAs have unique tissue distribution patterns | Manca et al. (34) |
| Cow milk | Ultracentrifugation | DLS, NTA, SEM, AFM, Western blot (CD63, CD81, Tsg101, Alix) | Milk exosomes can deliver chemotherapeutic drug paclitaxel and effectively inhibit tumor growth. | Agrawal et al. (35) |
| Cow colostrum | Density gradient centrifugation | Western blot (Alix, Tsg101) | Colostrum derived exosomes are enriched with proteins regulating the immune response and growth. | Samuel et al. (36) |
| Buffalo milk | Centrifugation Exiqon isolation kit | SEM, DLS | As delivery vehicle, exosome encapsulation enhances the stability, solubility and bioavailability of curcumin. | Vashisht et al. (37) |
OSCC, oral squamous cell carcinoma; TEM, transmission electron microscope; NTA, nanoparticle tracking analysis; NEC, necrotizing enterocolitis; ER, endoplasmic reticulum; DLS, dynamic light scattering; SEM, sanning electron microscopy; AFM, atomic force microscopy; AFM, atomic force microscopy.