A human tissue stem cell in which a gene of interest is introduced into a gene locus of a stem cell marker gene.

High mitochondrial ATP is a metabolic trait that confers hyper-proliferation, sternness, anchorage-independence, anti-oxidant capacity and multi-drug resistance in cancer cells. Under the present approach, intracellular ATP levels may be used as a metabolic biomarker to identify, separate, and purify an aggressive and hyper-proliferative cancer stem cell (“CSC”) phenotype. Further, ATP may be combined with other CSC markers, e.g., CD44 or ALDH-activity, to beneficially fractionate the CSC population into sub-populations. For example, ATP-high/ CD44-high CSC sub-populations showed twice the level of anchorage-independent growth compared to ATP-low/CD44-high CSC sub-populations. Also disclosed are complementary bioinformatic data that implicate mitochondrial ATP synthesis in stemness, metastasis, and the detection of circulating tumor cells (“CTCs”), and a five-member, ATP-related metastasis gene-signature (ABCA2, ATP5F1C, COX20, NDUFA2 and UQCRB). The gene signature of the present approach may be used to identify CSCs having a dramatic increase in cell migration and invasion in vitro capacity, as well as spontaneous metastasis in vivo. The present approach also provides a cellular platform for systematically targeting sternness, multi-drug resistance, and metastasis in cancer cells.

The present invention relates to a chemically defined medium for eukaryotic cell culture, comprising water, at least one carbon source, one or more vitamins, one or more salts, one or more growth factors, one or more fatty acids, one or more buffer components, selenium and one or more further trace elements and its use in the culture of cancer stem cells, in particular tumorsphere culture of cancer stem cells.

Disclosed are microcapsule compositions and methods for encapsulating living cells. The methods include a microencapsulation approach to isolate and culture high-quality stem cells, including human iPSCs, cancer stem cells, cardiac stem cells, and the like. The microencapsulation methods are inspired by the development of blastomeres into a blastocyst within the Zona pellucida of the human female reproductive system. The bioinspired methods include encapsulation of blastomere-like cell clusters in a Zona-like microcapsule including a miniaturized hyaluronic acid-rich core and a semipermeable hydrogel shell. The cell clusters are subsequently cultured to form highly pluripotent spheroids with improved cell quality, homogeneity, and viability. Methods of use of said microcapsules are also disclosed including therapeutic uses related to human iPSC-based personalized medicines.

The present application relates to a cell cryopreservation protective composition, the use of the composition, and a cell cryopreservation method. The cell cryopreservation protective composition comprises a zwitterionic molecule having the general formula R.sub.1—N.sup.+(CH.sub.3).sub.2—(CH.sub.2).sub.n—R.sub.2, wherein R.sub.1 is a linear or branched alkyl having 1 to 10 carbon atoms, and is optionally substituted with a substituent; R.sub.2 is any negatively charged group selected from the group consisting of —COO.sup.−, —SO.sub.4.sup.−, —SO.sub.3.sup.−, and (I); and R.sub.3 is a group selected from the group consisting of (methyl)acryloyloxyalkyl, alkyl and alkenyl; and the zwitterionic molecule having general formula R.sub.1—N.sup.+(CH.sub.3).sub.2—(CH.sub.2).sub.n—R.sub.2 is preferably a betaine compound. The cell cryopreservation protective composition can carry out cell cryopreservation in a non-toxic and efficient manner, and results in an extremely high post-thaw cell survival rate and does not require stepwise cryopreservation. After cell recovery, the cells can be used directly or after being slightly diluted. ##STR00001##

A method of characterizing a glioblastoma multiforme (GBM) stem cell (GSC), comprising culturing the GSC to provide a culture, contacting a first set of aliquots of the culture with individual compounds selected from a panel of compounds, identifying two or more of the selected compounds that cause more than a threshold level of cell death in the first set of aliquots, and characterizing the GSC as suitable for treatment with one or more combinations comprising the two or more identified compounds. A panel of chemical compounds, the compounds selected by a method comprising surgically resecting the tumor, culturing a GSC derived from GBM tissue derived from a GBM tumor, contacting aliquots thereof with individual compounds selected from a panel of compounds, and identifying two or more of the selected compounds that cause more than a threshold level of cell death in the aliquots, thereby identifying the compounds.

The present invention relates in part to methods for producing tissue-specific cells from patient samples, and to tissue-specific cells produced using these methods. Methods for reprogramming cells using RNA are disclosed. Therapeutics comprising cells produced using these methods are also disclosed.

Disclosed are compounds, methods, compositions, and kits that allow for treating cancer by, e.g., targeting cancer stem cells. In some embodiments, the cancer is colorectal cancer, gastric cancer, gastrointestinal stromal tumor, ovarian cancer, lung cancer, breast cancer, pancreatic cancer, prostate cancer, testicular cancer, or lymphoma. In some embodiments, the cancer is liver cancer, endometrial cancer, leukemia, or multiple myeloma. The compounds utilized in the disclosure are of Formula (0), (O′), and (I):

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The present invention provides cancer stem cell vaccines useful for treating or preventing a variety of tumors, as well as related methods of producing cancer stem cells and antigens thereof and producing vaccine adjuvants with enhanced activity for use with the stem cell vaccines.

Disclosed is a device for the culture of cells, which device is able to support and/or maintain the cells within an environment which mimics one or more in vivo environmental condition(s). Using these devices, cells can be cultured or maintained under conditions which ensure that the cells behave and respond substantially as they would in vivo. Further, the cells can be stimulated or exposed to exogenous agents (drugs and the like) and any response determined to be one which is indicative of an in vivo response.