3 research outputs found
Additional file 1: Figure S1-S16. of Cellular function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood post CRISPR genome editing
Function reinstitution of offspring red blood cells cloned from the sickle cell disease patient blood by a clinically practicable CRISPR/Cas9 method. (DOCX 4365 kb
Hand-Held and Integrated Single-Cell Pipettes
Successful single-cell isolation is a primary step for subsequent
chemical and biological analyses of single cells. Conventional single-cell
isolation methods often encounter operational complexity, limited
efficiency, deterioration of cell viability, incompetence in the isolation
of a single-cell into nanoliter liquid, and/or inability to select
single adherent cells with specific phenotypes. Here, we develop a
hand-held single-cell pipet (hSCP) that is rapid, operationally simple,
highly efficient, and inexpensive for unbiased isolation of single
viable suspended cells directly from submicroliter cell suspensions
into nanoliter droplets without the assistance of any additional equipment.
An integrated SCP (iSCP) has also been developed for selective isolation
of single suspended and adherent cells according to the fluorescence
imaging and morphological features. The isolated single cells can
be conveniently transferred into standard 96-/384-well plates, Petri
dishes, or vials for cloning, PCR, and other single-cell biochemical
assays
DNA Thioaptamer with Homing Specificity to Lymphoma Bone Marrow Involvement
Selective drug accumulation in the
malignant tissue is a prerequisite
for effective cancer treatment. However, most drug molecules and their
formulated particles are blocked en route to the destiny tissue due
to the existence of multiple biological and physical barriers including
the tumor microvessel endothelium. Since the endothelial cells on
the surface of the microvessel wall can be modulated by inflammatory
cytokines and chemokines secreted by the tumor or stromal cells, an
effective drug delivery approach is to enhance interaction between
the drug particles and the unique spectrum of surface proteins on
the tumor endothelium. In this study, we performed <i>in vivo</i> screening for thioaptamers that bind to the bone marrow endothelium
with specificity in a murine model of lymphoma with bone marrow involvement
(BMI). The R1 thioaptamer was isolated based on its high homing potency
to bones with BMI, and 40–60% less efficiency in accumulation
to healthy bones. In cell culture, R1 binds to human umbilical vein
endothelial cells (HUVEC) with a high affinity (<i>K</i><sub>d</sub> ≈ 3 nM), and the binding affinity can be further
enhanced when cells were treated with a mixture of lymphoma cell and
bone marrow cell conditioned media. Cellular uptake of R1 is through
clathrin-mediated endocytosis. Conjugating R1 on to the surface of
liposomal doxorubicin nanoparticles resulted in 2–3-fold increase
in drug accumulation in lymphoma BMI. Taking together, we have successfully
identified a thioaptamer that preferentially binds to the endothelium
of lymphoma BMI. It can serve as an affinity moiety for targeted delivery
of drug particles to the disease organ