Lymph node as a site for transplantation, organogenesis and function for multiple tissues and organs
AssigneeUniversity of Pittsburgh—Of the Commonwealth System of Higher Education
InventorEric Lagasse
Life of this patent
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Abstract
The present invention relates to methods and compositions for transplanting non-lymphoid tissues into lymphoid organs. It may be used to cultivate organ tissues including for the purpose of supplementing or reconstituting organ function. Tissues that may be propagated in this manner include but are not limited to lung, kidney, thyroid, intestine, and brain.
Claims12
1-17. (canceled)
A method of evaluating maturation of an ectopic graft epithelium, the method comprising:
- (a) obtaining RNA from the ectopic graft epithelium;
- (b) reverse transcribing the RNA from the ectopic graft epithelium to generate cDNA;
- and
- (c) performing a polymerase chain reaction (PCR) to quantify an amount of GM-CSF expressed in the ectopic graft epithelium;
- thereby evaluating the maturation of the ectopic graft epithelium.
The method of claim 1, wherein the RNA is obtained 21 weeks after the ectopic graft is administered to a subject.
The method of claim 1, wherein the PCR comprises about 25 reaction cycles.
The method of claim 1, wherein the PCR comprises a first primer comprising the nucleotide sequence of SEQ ID NO:
- 12 and a second primer comprising the nucleotide sequence of SEQ ID NO:
- 13.
The method of claim 1, further comprising comparing the amount of GM-CSF expressed in the ectopic graft epithelium to an amount of GM-CSF expressed in an E14.5/E.15.5 mouse thymus.
The method of claim 5, further comprising evaluating expression of keratin 8 in the ectopic graft epithelium and the E14.5/E.15.5 mouse thymus via immunofluorescence staining and fluorescence microscopy.
A nucleic acid comprising the nucleotide sequence of any one of SEQ ID NOs:
- 1-11.
A method, comprising:
- (i) backcrossing 129sv Fah−/− mice for more than eight generations with C57BL/6 mice, thereby generating a C57BL/6 Fah−/− mouse, and (ii) injecting a dye solution into a footpad of the C57BL/6 Fah−/− mouse.
The method of claim 8, wherein the dye solution comprises 3% evans blue solution.
The method of claim 8, wherein the dye solution is injected intradermally into the footpad of a hindlimb of the C57BL/6 Fah−/− mouse.
The method of claim 10, further comprising visualizing a popliteal lymph node in the hindlimb of the C57BL/6 Fah−/− mouse.
Dependencies are inferred from the claim text.