Methods for cryopreservation and resuscitation of follicles are provided by the present disclosure, relating to the technical field of follicular cryopreservation. According to the method for cryopreserving follicles provided by the present disclosure, follicles are firstly encapsulated with Alginate hydrogel, and incubated in protectant A and B. and moved into a protective solution A for incubation. Follicles encapsulated in hydrogel microcapsules are then loaded into straws and immediately submerged into liquid nitrogen for cryopreservation. Then, they are rapidly warmed by a 37? C. water bath combined with nano-warming technique. To verify the preservation effect, the warmed preantral follicles are further cultured in vitro in three dimensions.

Methods and materials for making and using apical-out ciliated organoids (e.g., apical-out airway organoids) are provided herein.

The present disclosure provides feeder hydrogel particles that can function to support the growth, proliferation, and/or activation of a target cell in culture. The present disclosure also provides methods of culturing target cells with feeder hydrogel particles.

This application relates to methods of generating cell/particle-free regions in a cellular/particle aggregate using label-free magnetic manipulation. For example, the method may include suspending cells/particles in a paramagnetic culture medium, the cells/particles being diamagnetic; seeding the cells/particles and the paramagnetic culture medium into microwells; and placing the microwells over an array of magnet pairs where each magnet pair creates a magnetic field gradient. The magnetic susceptibility difference and the magnetic field gradient drive the cells/particles towards a spaced region between magnets in the magnet pair having a lowest magnetic field strength to create a cell/particle aggregate with a cell-free/particle-free area where the cell/particle aggregate surrounds the cell-free/particle-free area.

Mutated genome silencing with endogenous RNAi-siRNA and miRNA with near total cellular apheresis with pulse flow apheresis system and EV-exosome-RNA molecular apheresis with sucrose density gradient continuous flow ultracentrifugation combined with array centrifuge for both 50S higher and 50S lower proteomics and genomics apheresis and their fractionated purification with immobilized Tim4-Fc protein Ca2+ magnetic beads affinity chromatography (ACG) and immobilized metal ACG is disclosed. It purifies normal cell derived and tumor cell derived EVs-exosomes, proteomics and subcellular particles. Tumor-specific endogenous siRNA is generated from mutated RNA containing pre-miRNA hairpin through RNA-induced silencing complex (RISC) composed of Dicer, dsRNA binding protein TRBP, and AGO2. Incubating purified RSIC with pre-let-7 hairpin generates siRNA. SiRNA is bonded with T-EVs and T-cells to silence its evasion from tumor immunity. While on radiation therapy or surgery, a patient's blood is continuously processed with above systems. It delivers combined online radiotherapy, and tumor-seeking adoptive extracorporeal chemo-immunotherapy.

Disclosed are methods, devices, and techniques useful for enhancing function of an organ or cellular graft through administration of light frequencies sufficient to enhance desired therapeutic activity. In one embodiment a bone marrow cellular population is treated with one or more wavelengths of low level laser irradiation at a sufficient energy to enhance engraftment, trophic, and regenerative activity in the corpus cavernosum. In another embodiment the recipient penile body is treated with one or more wavelengths of low level laser irradiation at a sufficient energy to enhance chemoattraction and growth factor secretion.

Therapies for treating erectile dysfunction using regenerative cells and therapeutic energy treatments are disclosed herein. Said therapeutic energy treatments can be selected from one or more of the following methods: electrical stimulation/electroacupuncture, low-level laser irradiation, and extracorporeal shockwave therapy. The combination treatments described herein are useful for restoring components of the penile anatomy that are associated with erectile dysfunction, in particular, nerves, blood vessels and/or smooth muscle cells.

The present invention provides methods of identifying and distinguishing different types of nerve cells in ex vivo cell culture, the method comprising the steps of: a) culturing somato-sensory nerve cells ex vivo, b) loading the nerve cells with a calcium, sodium, or voltage-sensitive indicator or expressing a genetically encoded calcium, sodium, or voltage-sensitive indicator, c) pulsing the nerve cells with an electrical train of bipolar square waves at two different voltages and two different frequencies; wherein the first voltage is 10 V/cm or less (low voltage) and the second voltage is between 12 and 20 V/cm (high voltage); and wherein the first frequency is 5 Hz or less (low frequency) and the second frequency is between 15 and 20 Hz (high frequency), and d) detecting activation of the nerve cell by measuring the changes in the signal intensity of the indicator, wherein low voltage and low frequency activation indicates a first type of cell and activation detected only at high voltage indicates a second type of cell.

A method for sterilising a platelet lysate in the liquid state comprising at least the endogenous growth factors TGF-beta 1, EGF, PDGF-AB, IGF-1, VEGF and bFGF. The method comprising freezing the liquid platelet lysate in order to obtain a frozen platelet lysate, and irradiating the frozen platelet lysate with ionising radiation in order to obtain a sterilised platelet lysate, the irradiation being adapted so as to preserve at least 80% of the concentration of at least one of the endogenous growth factors chosen from the group consisting of TGF-beta 1, EGF, PDGF-AB, IGF-1 and VEGF.

A simple and efficient method for separating migrasomes from macrophages is provided. The method separates migrasomes having a diameter range of 0.5 micrometers (?m) to 3 ?m by intercepting through a filter and eluting through reverse filtration successfully. The separated migrasome has a vesicle-liked structure and wrinkles on its surface, and the separated migrasome has a diameter over 500 nanometers (nm). The separated migrasomes express their characteristic proteins PIGK, EOGT, and TSPAN4, but do not express specific markers TSG101 and ALIX of EVs, indicating that the separated migrasomes are a unique type of vesicles distinct from extracellular vesicles (EVs). The integrity of ribonucleic acids (RNA) carried by the migrasomes is not affected. The method for separating migrasomes from macrophages has the characteristics of simplicity, high efficiency, good controllability, good repeatability, and low cost, and large special equipment is not needed.