FOETAL POLYMIX OF MESENCHYMAL STEM CELLS UNDER HYPOXIC CONDITIONS FOR THE TREATMENT OF CLINICAL DISORDERS AND DISEASES

1 - 25 . (canceled)

26 . A method making a foetal polymix composition, the method comprising:

isolating mesenchymal stem cells (MSCs) from two separate sources, wherein the two sources are placenta and umbilical cord tissue;

enriching the MSCs from each source separately; and

co-culturing the enriched MSCs from each source together in a culture medium under hypoxic conditions, wherein the hypoxic conditions comprise incubation in a carbon dioxide incubator at 37° C. with 1% oxygen concentration, and with a remaining concentration containing carbon dioxide and nitrogen,

wherein the MSCs from placenta are isolated and enriched by a process comprising of:

collecting a chorionic trophoblast from the placenta;

washing the chorionic trophoblast with a sterile phosphate buffer solution (PBS);

selecting an area from the chorionic trophoblast that contains chorionic wherein the area is minced into a plurality of pieces;

adding collagenase and dispase to a container containing minced chorionic trophoblast for digestion;

incubating the container;

adding fetal bovine serum to the container to inhibit activity of collagenase and dispase;

filtering a mixture in the container; centrifuging a filtrate from the mixture to obtain a pellet;

re-suspending the pellet in another PBS to obtain a re-suspended pellet layer;

adding the re-suspended pellet layer on top of a ficoU density gradient medium to obtain a mixture;

centrifuging the mixture to obtain cells from a middle band;

washing the cells with PBS to obtain mesenchymal stem cells;

re-suspending the mesenchymal stem cells in Dulbecco's Modified Eagle's Medium (DMEM) for culturing;

seeding isolated mesenchymal stem cells from DMEM into a first culture vessel; and

incubating the seeded isolated mesenchymal stem cells until the seeded isolated mesenchymal stem cells reach 70-80% confluence,

wherein the MSCs from an umbilical cord are isolated and enriched by a process comprising of:

subjecting a processed umbilical cord to an enzymatic digestion with collagenase and hyaluronidase to form a mixture;

incubating the mixture containing the processed umbilical cord, collagenase, and hyaluronidase with gentle agitation;

adding trypsin to the agitated mixture and then incubating to obtain a digested suspension;

diluting the digested suspension with phosphate buffer solution (PBS) to obtain a diluted suspension;

filtering the diluted suspension to obtain a single cell suspension;

centrifuging the single cell suspension to obtain a pellet;

re-suspending the pellet in a washing solution comprising albumin, hydroxyethyl starch (HES), and citrate-phosphate-dextrose-adenine anticoagulant;

isolating cells containing MSCs from the washing solution;

seeding isolated MSCs with DMEM into a second culture vessel; and

incubating the seeded isolated MSCs until the seeded isolated MSCs reach 70-80% confluence.

27 . The method of claim 26 , further comprising wherein the foetal polymix composition is formulated into an injectable composition.

28 . The method of claim 26 , further comprising wherein the foetal polymix composition is formulated to be suitable for administering via an intravenous route, an intrathecal route, an intra-articular route, an intramuscular route, a derma roller, a mesogun, a local route, a periorbital route, an intravitreal route, or a retrobular route.

29 . The method of claim 26 , further comprising wherein the foetal polymix composition is formulated to be suitable for treating disorders and diseases.

30 . A method making a foetal polymix composition, the method comprising:

isolating mesenchymal stem cells (MSCs) from two separate sources, wherein the two sources are placenta and umbilical cord tissue;

enriching the MSCs from each source separately; and

co-culturing the enriched MSCs from each source together in a culture medium under hypoxic conditions, wherein the hypoxic conditions comprise incubation in a carbon dioxide incubator at 37° C. with 1% oxygen concentration, and with a remaining concentration containing carbon dioxide and nitrogen,

wherein the MSCs from placenta are isolated and enriched by a process comprising of:

collecting a fetal decidua's placental tissue from placenta;

washing the fetal decidua's placental tissue with a phosphate buffer solution (PBS);

digesting the fetal decidua's placental tissue with collagenase;

separating cells by centrifuging a mixture of the fetal decidua's placental tissue and collagenase over a Ficoll-Hypaque separation medium to obtain separated cells;

re-suspending the separated cells for culture in L-Dulbecco's Modified Eagle's Medium (LDMEM) supplemented with fetal bovine serum, basic fibroblast growth factor, L-glutamine, and at least one antibiotic;

seeding cells from the LDMEM into a first culture vessel;

removing non-adherent cells from the first culture vessel;

harvesting cells from the first culture vessel by adding trypsin and ethylenediaminetetraacetic acid (EDTA); and

passaging the cells,

wherein the MSCs from an umbilical cord are isolated and enriched by a process comprising of:

subjecting a processed umbilical cord to an enzymatic digestion with collagenase and hyaluronidase to form a mixture;

incubating the mixture containing the processed umbilical cord, collagenase, and hyaluronidase with gentle agitation;

adding trypsin to the agitated mixture and then incubating to obtain a digested suspension;

diluting the digested suspension with phosphate buffer solution (PBS) to obtain a diluted suspension;

filtering the diluted suspension to obtain a single cell suspension;

centrifuging the single cell suspension to obtain a pellet;

re-suspending the pellet in a washing solution comprising albumin, hydroxyethyl starch (HES), and citrate-phosphate-dextrose-adenine anticoagulant;

isolating cells containing MSCs from the washing solution;

seeding isolated MSCs with DMEM into a second culture vessel; and

incubating the seeded isolated MSCs until the seeded isolated MSCs reach 70-80% confluence.

31 . The method of claim 30 , further comprising wherein the foetal polymix composition is formulated into an injectable composition.

32 . The method of claim 30 , further comprising wherein the foetal polymix composition is formulated to be suitable for administering via an intravenous route, an intrathecal route, an intra-articular route, an intramuscular route, a derma roller, a mesogun, a local route, a periorbital route, an intravitreal route, or a retrobular route.

33 . The method of claim 30 , further comprising wherein the foetal polymix composition is formulated to be suitable for treating disorders and diseases.

34 . A method making a foetal polymix composition, the method comprising:

isolating mesenchymal stem cells (MSCs) from two separate sources, wherein the two sources are placenta and umbilical cord tissue;

enriching the MSCs from each source separately; and

co-culturing the enriched MSCs from each source together in a culture medium under hypoxic conditions, wherein the hypoxic conditions comprise incubation in a carbon dioxide incubator at 37° C. with 1% oxygen concentration, and with a remaining concentration containing carbon dioxide and nitrogen,

wherein the MSCs from placenta are isolated and enriched by a process comprising of:

collecting a chorion from the placenta;

mincing the chorion to obtain a minced chorion;

centrifuging the minced chorion to obtain a first pellet;

haemolysing the first pellet with a red blood cell lysis buffer;

centrifuging a mixture comprising the first pellet and the red blood cell lysis buffer to obtain a second pellet;

incubating the second pellet with a trypsin-ethylenediaminetetraacetic acid (EDTA) solution to obtain a trypsin-EDTA solution treated pellet;

re-suspending the trypsin-EDTA solution treated pellet in a minimum essential medium (MEM) or a fetal bovine serum to obtain a re-suspended mixture;

centrifuging the re-suspended mixture to obtain a third pellet; and

culturing, in a first culture vessel, the third pellet comprising mesenchymal stem cells in a minimum essential medium (MEM) supplemented with fetal calf serum, L-ascorbate-2-phosphate, L-Glutamine, an antibiotic, fungizone, and tylosin,

wherein the MSCs from an umbilical cord are isolated and enriched by a process comprising of:

subjecting a processed umbilical cord to an enzymatic digestion with collagenase and hyaluronidase to form a mixture;

incubating the mixture containing the processed umbilical cord, collagenase, and hyaluronidase with gentle agitation;

adding trypsin to the agitated mixture and then incubating to obtain a digested suspension;

diluting the digested suspension with phosphate buffer solution (PBS) to obtain a diluted suspension;

filtering the diluted suspension to obtain a single cell suspension;

centrifuging the single cell suspension to obtain a pellet;

re-suspending the pellet in a washing solution comprising albumin, hydroxyethyl starch (HES), and citrate-phosphate-dextrose-adenine anticoagulant;

isolating cells containing MSCs from the washing solution;

seeding isolated MSCs with DMEM into a second culture vessel; and

incubating the seeded isolated MSCs until the seeded isolated MSCs reach 70-80% confluence.

35 . The method of claim 34 , further comprising wherein the foetal polymix composition is formulated into an injectable composition.

36 . The method of claim 34 , further comprising wherein the foetal polymix composition is formulated to be suitable for administering via an intravenous route, an intrathecal route, an intra-articular route, an intramuscular route, a derma roller, a mesogun, a local route, a periorbital route, an intravitreal route, or a retrobular route.

37 . The method of claim 34 , further comprising wherein the foetal polymix composition is formulated to be suitable for treating disorders and diseases.