Microfluidic devices and methods for co-encapsulation of a cell and a controlled release particle in one droplet are escribed herein. The devices and methods utilize laminar flow, high shear liquid-liquid interfaces, hydrodynamic vortices, and/or acoustic focusing to increase co-encapsulation efficiency. The precise variation of the droplets microenvironment is enabled by the controlled release particle co-encapsulated with the single cell in each droplet. This capability, coupled with established detection methods, provides an important tool for precise, single cell analysis.

The present disclosure describes methods for intracellular electromanipulation of proteins using nanosecond pulsed electric fields (nsPEFs). The nsPEFs have effects on proteins in addition to permeabilizing cellular membranes. The nsPEFs induce a Ca.sup.2+-dependent dissipation of the mitochondria membrane potential (ΔΨm), which is enhanced when high frequency components are present in fast rise-fall waveforms. Ca.sup.2+ is shown to have little or no effect on propidium iodide uptake as a measure of plasma membrane poration and consequently intracellular membranes. Since Ca.sup.2+-regulated events are mediated by proteins, actions of nsPEFs on proteins that regulate and/or affect the mitochondria membrane potential are possible. Given that nsPEF-induced dissipation of ΔΨm was more effective when high frequency components were present in fast rise time waveforms, the effects on proteins are due to these high frequency components. These results present direct evidence that nsPEFs affect proteins and their functions by affecting their structure.

Provided herein are models comprising a tissue-like assembly of cells tissues or organoid bodies cultured in the presence of a pulsating alternating ionic magnetic resonance field. The cells, tissues or organoid bodies are introduced into a culture unit comprising growth and nutrient modules in which the gravity vector of the growth unit is continually randomized and cultured in the presence of the alternating ionic magnetic resonance field.

The present invention is directed generally to systems for making host cells with artificial endosymbionts. In an embodiment, the systems of the invention comprise a host cell, an artificial endosymbiont, a means for introducing the artificial endosymbiont to the host cell, a means for separating the host cells with artificial endosymbionts from free artificial endosymbionts, and a means for detecting artificial endosymbionts in host cells.

The disclosure provided herein relates generally to mesenchymal-like stem cells “hES-T-MiSC” or “T-MSC” and the method of producing the stem cells. The method comprises culturing embryonic stem cells under conditions that the embryonic stem cells develop through an intermediate differentiation of trophoblasts, and culturing the differentiated trophoblasts to hES-T-MSC or T-MSC, T-MSC derived cells and cell lineages “T-MSC-DL” are also described. Disclosed also herein are solutions and pharmaceutical compositions comprising the T-MSC and/or T-MSC-DL, methods of making the T-MSC and T-MSC-DL, and methods of using the T-MSC and T-MSC-DL for treatment and prevention of diseases, specifically, T-MSC and T-MSC-DL are used as immunosuppressive agents to treat multiple sclerosis and autoimmune diseases.

A systemic human repair and enhancement system that provides stem cell therapy utilizing an apparatus and methods for administering a treatment schedule of non-invasive and drug-free narrow and specific 0.6180 Hz frequency continuous sine wave therapy to the patient's cells. The frequency therapy may be in-vivo and in-vitro applications. The frequency therapy stimulates systemic stem cell production, particularly at a tissue sites having loss of function due to a condition, aging, damage, and or disease. The frequency therapy system encompasses a narrow and specific ultra-low continuous sine wave frequency stimulation therapy that produces one or more of the enhanced cell production, release, viability, proliferation, migration and or engraftment of the human patient's own genetically compatible stem cells, and the therapy enhances the stem cell's secretions thereby safely enhancing the secretion's therapeutic effects during stem cell therapy and simultaneously systemically enhancing the patient's pre-existing cells and tissues.

The present disclosure provides metal-containing (MC) semiconducting (SC) Pdots (MC-SC-Pdots) with beneficial functionalities in both cellular imaging and manipulation, among other applications. The Pdots comprise at least one nanoparticle comprising at least one metal, and a semiconducting polymer associated with the nanoparticle.

Methods of making and using a magnetic ECM are disclosed. The ECM comprises positively and negatively charged nanoparticles, wherein one of said nanoparticles contains a magnetically responsive element. When the magnetic ECM is seeded with cells, the cells will be magnetized and can be levitated for 3-D cell culture.

A method of manufacturing and/or using a scaffold assembly for stem cell culture and tissue engineering applications is disclosed. The scaffold at least partially mimics a native biological environment by providing biochemical, topographical, mechanical and electrical cues by using an electroactive material. The assembly includes at least one layer of substantially aligned, electrospun polymer fiber having an operative connection for individual voltage application. A method of cell tissue engineering and/or stem cell differentiation that uses the assembly seeded with a sample of cells suspended in cell culture media, incubates and applies voltage to one or more layers, and thus produces cells and/or a tissue construct. In another aspect, the invention provides a method of manufacturing the assembly including the steps of providing a first pre-electroded substrate surface; electrospinning a first substantially aligned polymer fiber layer onto the first surface; providing a second pre-electroded substrate surface; electrospinning a second substantially aligned polymer fiber layer onto the second surface; and, retaining together the layered surfaces with a clamp and/or an adhesive compound.

A method for producing exosomes comprises steps of: providing ultrasound stimulation directly or indirectly to cells; culturing a mixture of the cells and a medium for a predetermined time; and isolating exosomes from the mixture, wherein providing the stimulation directly to the cells comprises applying ultrasound stimulation to the medium containing the cells, and the providing the stimulation indirectly to the cells comprises applying ultrasound stimulation to the medium not containing the cells and then mixing the medium and the cells. This method for producing exosomes makes it possible to obtain exosomes having a hair regeneration effect not only from stem cells and progenitor cells that are difficult to isolate and multiply, but also from somatic cells that may be easily obtained and maintained, in high yield within a short time by ultrasound treatment that is a simple process.