The present application discloses a method for preventing or retarding degeneration of intervertebral disc at an intervertebral disc defect site, which includes injecting a mammalian connective tissue cell into the intervertebral disc defect site.

This application relates to a method of making a cell construct, comprising a) plating a plurality of cells on a substantially flat surface; b) growing the plurality of cells to at least 80% confluent to form a cell sheet with intercellular linkages; c) applying a culture medium having a pH of about 5 to about 6.8 to the cell sheet; d) replacing the culture medium of step c) with a culture medium having a pH of about 7.5 to about 8.5; and e) replacing the culture medium of step d) with a culture medium having a pH of about 7 to about 7.7, to obtain a substantially planar untethered cell sheet. Also provided is a cell construct formed according to the method and uses thereof.

This application relates to a method of making a cell construct, comprising a) plating a plurality of cells on a substantially flat surface; b) growing the plurality of cells to at least 80% confluent to form a cell sheet with intercellular linkages; c) applying a culture medium having a pH of about 5 to about 6.8 to the cell sheet; d) replacing the culture medium of step c) with a culture medium having a pH of about 7.5 to about 8.5; and e) replacing the culture medium of step d) with a culture medium having a pH of about 7 to about 7.7, to obtain a substantially planar untethered cell sheet. Also provided is a cell construct formed according to the method and uses thereof.

Preparation of a product comprising stromal vascular traction cells includes washing human biological material comprising adipose in a container apparatus having an internal filter, which divides an internal containment volume of the container apparatus into a tissue retention volume on one side of the filter and a filtrate volume on an opposite side of the filter, and a mixing device with at least one rotatable mixing member disposed in the tissue retention volume. The washing includes operation of the mixing device to rotate the mixing member through the human biological material within the tissue retention volume, and the washing is followed by digesting washed material within the internal containment volume with added enzyme, centrifuging of the container apparatus to prepare a centrifuged pellet in the filtrate volume, selectively removing material of the pellet and preparing a product with a mixture of stromal vascular fraction cells of removed pellet material and an aqueous suspension liquid.

Preparation of a product comprising stromal vascular traction cells includes washing human biological material comprising adipose in a container apparatus having an internal filter, which divides an internal containment volume of the container apparatus into a tissue retention volume on one side of the filter and a filtrate volume on an opposite side of the filter, and a mixing device with at least one rotatable mixing member disposed in the tissue retention volume. The washing includes operation of the mixing device to rotate the mixing member through the human biological material within the tissue retention volume, and the washing is followed by digesting washed material within the internal containment volume with added enzyme, centrifuging of the container apparatus to prepare a centrifuged pellet in the filtrate volume, selectively removing material of the pellet and preparing a product with a mixture of stromal vascular fraction cells of removed pellet material and an aqueous suspension liquid.

Method for preparing and administering human pluripotent stem cells includes preparing human adipose stem cells from a human; epigenetically modifying the human adipose stem cells (hADSC) to yield directly-generated human pluripotent stem cells (dgHPSC); and engineering the dgHPSC to secrete a therapeutic level of insulin. Preparing the human adipose stem cells includes obtaining a lipoaspirate from a human and preparing adipose stem cells from the lipoaspirate. Epigenetically modifying the hADSC includes inducing the hADSC to yield the dgHPSC. Engineering the dgHPSC includes transducing a human estrogen-related receptor gamma (ERRγ) gene into the dgHPSC. Engineering the dgHPSC further includes transducing a human INS gene into the dgHPSC. The dgHPSC transduced with the ERRγ gene and the human INS gene secrete a higher level of insulin compared to the dgHPSC transduced with the ERRγ gene. The engineered dgHPDC are introduced into a human.

Methods of producing human stem cells are disclosed for parthenogenetically activating human oocytes by manipulation of O.sub.2 tension, including manipulation of Ca.sup.2+ under high O.sub.2 tension and contacting oocytes with serine threonine kinase inhibitors under low O.sub.2 tension, isolating inner cell masses (ICMs) from the activated oocytes, and culturing the cells of the isolated ICMs under high O.sub.2 tension. Moreover, methods are described for the production of stems cells from activated oocytes in the absence of non-human animal products, including the use of human feeder cells/products for culturing ICM/stem cells. Stem cells produced by the disclosed methods are also described.

Methods of producing human stem cells are disclosed for parthenogenetically activating human oocytes by manipulation of O.sub.2 tension, including manipulation of Ca.sup.2+ under high O.sub.2 tension and contacting oocytes with serine threonine kinase inhibitors under low O.sub.2 tension, isolating inner cell masses (ICMs) from the activated oocytes, and culturing the cells of the isolated ICMs under high O.sub.2 tension. Moreover, methods are described for the production of stems cells from activated oocytes in the absence of non-human animal products, including the use of human feeder cells/products for culturing ICM/stem cells. Stem cells produced by the disclosed methods are also described.

A composition for treating a patient with a tissue disease or malformity has a composition containing amniotic fluid. The amniotic fluid has a quantity of gender specific amniotic fluid based on a gender of a fetal source. A method of treating a patient with a tissue disease or malformity comprises the steps of: identifying the tissue region to be treated and selecting a location to apply either topically or by injection or inhalation a composition containing amniotic fluid; selecting the composition containing amniotic fluid wherein the amniotic fluid has a quantity of gender specific amniotic fluid based on a gender of a fetal source allowing more specific targeted growth factors to be used for specific disease processes; and applying or injecting the composition at or into the selected location.

A composition for treating a patient with a tissue disease or malformity has a composition containing amniotic fluid. The amniotic fluid has a quantity of gender specific amniotic fluid based on a gender of a fetal source. A method of treating a patient with a tissue disease or malformity comprises the steps of: identifying the tissue region to be treated and selecting a location to apply either topically or by injection or inhalation a composition containing amniotic fluid; selecting the composition containing amniotic fluid wherein the amniotic fluid has a quantity of gender specific amniotic fluid based on a gender of a fetal source allowing more specific targeted growth factors to be used for specific disease processes; and applying or injecting the composition at or into the selected location.