There is disclosed an improved, safer and commercially efficient process for developing genetically engineered cells. More specifically, there is disclosed a process comprises introducing a donor DNA construct, a guide RNA, and an RNA-guided nuclease with the host cells to he transfected; and introducing the three components into the host cell. There is further disclosed a donor DNA construct designed for inserting a CAR (chimeric antigen receptor) into a defined genomic site of a host cell. Further, the present disclosure provides a host cell transfected with a CAR that lacks viral vectors that can present a safety concern. The disclosure provides for more efficient and more cost-effective process for engineering T cells to express CAR constructs.

Provided are a composition containing (−)-Blebbistain and/or para-Blebbistain, a kit comprising the composition, and a cardiomyocyte culture method using the composition as a culture medium. Also provided is a use of (−)-Blebbistain and/or para-Blebbistain in preparation of a cardiomyocyte isolation reagent or cardiomyocyte culture reagent.

Solutions to the problem of the invention are a method for selectively killing or removing a senescent cell, substance identification, and a method for purifying a senescent cell. Specifically, the invention includes an agent for removing a senescent cell, which is a drug for removing an in vivo senescent cell, the agent containing an inhibitor for glutaminase as an active ingredient, and a pharmaceutical composition containing the agent. The invention further includes a method for preparing a senescent cell, including the following steps (a) to (c): (a) synchronizing a cell with the G2 phase; (b) activating an intracellular p53 protein in the cell synchronized with the G2 phase; and (c) inhibiting polo-like kinase 1 (PLK1) activity in the cell treated in the step (b).

Provided is a method for culturing immature oocytes. The method can promote in vitro maturation of the immature oocytes, and specifically comprises using follicular cells and a culture medium for culturing same. The culture medium for culturing the follicular cells contains CNP or variants thereof or analogues thereof and an HDAC (histone deacetylase) inhibitor. Also provided are the in vitro maturation culture medium containing CNP or variants thereof or analogues thereof and the HDAC inhibitor, and related compositions thereof, and the use of the above medium, culture medium and compositions in the promotion of in vitro maturation of the immature oocytes.

The present invention provides a method for inducing an inversion of normal blood coagulation factor VIII (F8) gene, a method for correcting an inversion of blood coagulation factor VIII gene in which the inversion has occurred, and a Hemophilia A patient-derived induced pluripotent stem cell in which the inversion is corrected, constructed using the same. The method of the present invention effectively reproduces the inversion of intron 1 and intron 22 of the F8 gene, which is responsible for the majority of severe hemophilia A, and thereby may be effectively used for studying the development mechanism of hemophilia A and as a research tool for screening therapeutic agents. The inversion-corrected induced pluripotent stem cell constructed according the method of the present invention enables an efficient and fundamental treatment for hemophilia A by restoring a genotype in which mutation has occurred to a wild type-like state, without limitation via normal gene or protein delivery.

Provided herein are methods for the production of tissue resident memory-like T cells by the combination of hypoxia and TGFβ. Further provided herein are methods of using the tissue resident memory T cells as adoptive cell therapy.

Provided herein are methods of generating MDSCs ex vivo. The methods include culturing blood cells with lactoferrin.

Provided are chemical inducers of pluripotency (CIP) which include glycogen synthase kinase inhibitors, TGFβ receptor inhibitors, cyclic AMP agonists and S-adenosylhomocysteine hydrolase (SAH) inhibitors or histone acetylators. A method of inducing pluripotency in a partially or completely differentiated cell by using such chemical inducers of pluripotency is also provided. The method includes: (i) contacting a cell with the CIPs for a sufficient period of time to result in reprograming the cell into a pluripotent stem cell having ESC-like characteristics (CiPSC). Isolated chemically induced pluripotent stem cells (CiPSCs) and their progeny, produced by inducing differentiation of the CiPSCs, can be used in a number of applications, including but not limited to cell therapy and tissue engineering.

A culture medium and a culture method for neural cells, particularly relating to a composition comprises a SMAD signaling pathway inhibitor, a SHH signaling pathway agonist, a Wnt signaling pathway agonist and a Myosin II ATPase inhibitor, optionally, it further comprises a ROCK inhibitor; or, the composition consists of the SMAD signaling pathway inhibitor, the SHH signaling pathway agonist, the Wnt signaling pathway agonist, the Myosin II ATPase inhibitor, and optionally the ROCK inhibitor. The same batch of neural cells, such as midbrain dopaminergic progenitors, can be efficiently obtained by using the culture method and culture medium designed on the basis of the composition.

The present invention relates to a medium for direct differentiation of embryonic stem cell-derived mesenchymal stem cells, a method of preparing mesenchymal stem cells by using same, mesenchymal stem cells prepared thereby, and a cell therapy product comprising the same mesenchymal stem cells. In a medium composition and a method according to an embodiment, mesenchymal stem cells may be prepared at high yield within a short period of time. In addition, the method is simple in preparation procedure because of the absence of an embryoid body formation step and allows homogeneous cells to be prepared, thus advantageously providing a cell therapy product within a reduce period of time, compared to conventional methods.