Methods of producing stem cell conditioned media to treat mammalian injuries or insults. In at least one embodiment of a method for isolating non-endothelial adipocyte-depleted stromal cells of the present disclosure, the method comprises, comprising dissociating subcutaneous adipose tissue isolated from a mammal into a cell suspension, removing adipocytes from said cell suspension, resulting in a non-endothelial adipocyte-depleted cell suspension, and culturing the non-endothelial adipocyte-depleted cell suspension in a media containing growth factors VEGF, bFGF, EGF, and IGF, such that a mixed population of cells comprising a first population of further differentiated non-endothelial adipocyte-depleted CD34+/VE-cadherin− cells and a second population of further differentiated non-endothelial adipocyte-depleted CD34+/VE-cadherin+ cells are obtained and expanded.

The invention provides a method of producing a connective tissue graft suitable for correcting a connective tissue defect, comprising determining the size and shape of a tissue defect, obtaining a fat tissue from a patient modelled to fit the size and shape of the tissue defect, contacting the fat tissue with one or more connective tissue specific growth or differentiation factors; and kits for such a method.

An object of the present invention is to provide a method capable of inexpensively and conveniently preparing cells having pluripotency and a very low risk of tumorigenic transformation. The cells having pluripotency and a very low risk of tumorigenic transformation can be prepared by suspension-culturing mammalian mesenchymal stem cells such as human mesenchymal stem cells from bone marrow (hMSC-BM) and human adipose tissue-derived mesenchymal stem cells (hAT-MSC) (also referred to as “human adipose-derived stem cells [hADSC]”), 7 types of human adherent mature cells (human hepatocyte cells [hHEP cells], human umbilical vein endothelial cells [HUVEC cells], human dermal lymphatic microvascular endothelial cells [HMVEC cells], human epidermal keratinocyte cells [NHEK cells], human bronchial epithelial cells [NHBE cells], human melanocyte cells [NHEM cells], and human smooth muscle cells [UASMC cells]), and 3 types of human adherent precursor cells (human dermal fibroblast cells [NHDF cells], human skeletal muscle myoblast cells [HSMM cells], and human osteoblast cells [NHOst cells]) to form cell masses (spheroids).

The present invention addresses the problem of providing a method for obtaining Schwann cells directly (by direct reprogramming) without passing through pluripotent stem cells, such as ES cells or iPS cells. As a means for solving this problem, the present invention provides a method for preparing Schwann cells that includes a step of introducing into somatic cells of a mammal at least one gene selected from the group consisting of SOX10 genes and KROX20 genes, or an expression product thereof.

A serum-free complete medium for inducing differentiation of a mesenchymal stem cell to a corneal epithelial cell in the field of differentiation induction of stem cells, prepared by the following method: uniformly mixing the serum-free complete medium, containing 5-10 μmol of resveratrol, 2-4 μmol of icariin, 1-3 nmol of aspirin, 1-3 nmol of parathyroid hormone, 5-10 nmol of hydrocortisone, 1-3 mg of rapamycin, 2-10 μg of testosterone, 2-10 μg of EPO, 2-10 μg of LIF and the balance of a corneal epithelial cell serum-free medium in per 1 L; and then performing sterilization by filtration. The disclosure uses resveratrol and icariin in combination with aspirin, parathyroid hormone, hydrocortisone, rapamycin, testosterone and growth factors to cooperatively induce directional differentiation, uses nontoxic induction components, is high in induction efficiency and short in induction time, and achieves high induced corneal epithelial cell activity, no cell transplantation rejection, no ethical problem and high safety.

Methods for the efficient isolation and use of pluripotent adipose-derived stem cells (PASCs) are provided. In certain embodiments the methods involve providing an adipose tissue sample from which the stromal vascular fraction is co-cultured with the adipocyte fraction. PASCs can be isolated with a high degree of purification without requiring an additional cell enrichment process (e.g. cell sorting). PASCs and their conditioned media can be used for tissue regeneration within hours of harvesting the adipose tissue, and without requiring cell expansion. PASCs can grow as floating individual cells, as clusters of cells, or attached to surface(s) of the culture vessel. PASCs do not produce teratomas in vivo, nor do they induce immunorejection upon transplantation, and they achieve a high efficiency in grafting. The cells and compositions can be used for cell therapy and to screen new drugs.

The presently disclosed subject matter relates generally to the delivery of electrical stimuli via cell-penetrating nanoelectrodes. Such electrical stimuli leads to differentiation of cells, including but not limited to adipose derived stem cells, to neural lineage, specifically to neural cells.

The present invention has developed methods and compositions utilizing the cell surface CD107a (LAMP-1) as a marker that divides bone-forming and fat-forming progenitor/stem cells within human adipose tissue. The present invention is able to partition stromal progenitors for improved bone and soft tissue engineering and therapies.

The present invention relates to a cell differentiation medium composition, a high secretion insulin-producing cells and a preparation method thereof. The high secretion insulin-producing cells obtained by using the cell differentiation medium composition to induce stem cell differentiated under specific conditions can secrete a large amount of insulin in a short time, and when the high-secreting insulin-producing cells are transplanted into the human body, they are not easy to be swallowed by macrophages, which can improve the survival rate of the insulin-producing cells and prolong the time of insulin secretion thereby.

Provided herein are methods and systems for bio-printing of three-dimensional organs and organoids. Also provided herein are bio-printed three-dimensional organs and organoids for use in the generation and/or the assessment of immunological products and/or immune responses. Also provided herein are methods and system for bio-printing three-dimensional matrices.