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US 12,150,962 B2Granted
Method for producing extracellular vesicles comprising fetus-derived primal immunoglobulin
AssigneeStemmedicare Co., Ltd.
InventorJang Ho Lee
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Abstract
A fetal stem cell-derived extracellular vesicle has immune tolerance properties. The extracellular vesicle containing a fetal stem cell-derived primal immunoglobulin contains various natural antibodies and complement proteins which can immediately respond to foreign infectious agents such as viruses, bacteria, pathogens, etc., and thus can effectively treat and prevent infectious diseases through an enhanced innate immune system. Methods are disclosed for preparing the fetal stem cell-derived extracellular vesicle.
Claims6
An extracellular vesicle derived from human fetal stem cells, the extracellular vesicle comprising:
- immunoglobulin M, immunoglobulin A, immunoglobulin D, immunoglobulin G, immunoglobulin G3, and immunoglobulin E,
- wherein the extracellular vesicle further contains HLA-G proteins,
- wherein the extracellular vesicles are prepared by:
- (a) obtaining human trophoblast-derived extracellular vesicles from the human trophoblast expressing and secreting HLA-G;
- (b) obtaining extracellular vesicles from the hypoxic co-culture of human hematopoietic stem cells and human umbilical cord blood-derived mesenchymal stem cells;
- (c) inoculating the human fetal stem cells derived from amniotic fluid obtained during amniocentesis around 15 weeks of pregnancy into a matrix gel comprising hyaluronic acid, and subculturing in a serum-free medium containing the trophoblast-derived extracellular vesicles of step (a) and the extracellular vesicles of step (b);
- (d) inoculating the subcultured fetal stem cells of step (c) into a culture plate and culturing in a serum-free medium, so as to obtain a culture supernatant;
- and
- (e) filtering the culture supernatant and isolating the extracellular vesicles derived from human fetal stem cells,
- wherein the trophoblast-derived extracellular vesicles of step (a) are prepared by:
- (a-1) obtaining extracellular vesicles from the co-culture of human amniotic membrane-derived mesenchymal stem cells and amniotic fluid-derived mesenchymal stem cells;
- (a-2) inoculating human trophoblast cells into a matrix gel containing hyaluronic acid in the presence of the extracellular vesicles of step (a-1);
- and
- (a-3) culturing the trophoblast cells of step (a-2),
- wherein the extracellular vesicles obtained from the hypoxic co-culture of human hematopoietic stem cells and umbilical cord blood-derived mesenchymal stem cells of step (b) are prepared by:
- (b-1) inoculating human hematopoietic stem cells and umbilical cord blood-derived mesenchymal stem cells in upper and lower wells of a co-culture plate, respectively;
- (b-2) co-culturing in a serum-free medium under 1 to 5% O2 to obtain a culture supernatant;
- and
- (b-3) filtering the culture supernatant and isolating the extracellular vesicles.
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