Abstract
In vivo inoculation of cells such as tumor cells and induced pluripotent stem (iPS)/embryonic stem (ES) cells into immunocompromised mice has been considered as a powerful technique to evaluate their potential to proliferate or differentiate into various cell types originating from three germ cell layers. Subcutaneous grafting and grafting under the kidney capsule have been widely used for this purpose, but there are some demerits such as the requirement of a large number of tumor cells for inoculation and frequent failure of tumorigenesis. Therefore, grafting into other sites has been explored, including intratesticular or intramuscular grafting as well as grafting into the cochleae, liver, or salivary glands. In this study, we found that intrapancreatic parenchymal injection of cells is useful for allowing a small number of cells (~15 × 103 cells or ~30 cell clumps μL−1·site−1) to proliferate and sometimes differentiate into various types of cells. It requires only surgical exposure of the pancreas over the dorsal skin and subsequent injection of cells towards the pancreatic parenchyma under dissecting microscope-based observation using a mouthpiece-controlled glass micropipette. We now name this technology “intrapancreatic parenchymal cell transplantation (IPPCT)”, which will be useful, especially when only a small number of cells or colonies are available.
Highlights
Induced pluripotent stem/embryonic stem (ES) cells have the potential to differentiate into fully differentiated cells originating from three germ layers when they are placed under differentiation-inducing conditions [1,2]
The exposed pancreas of an anesthetized nude mouse was subjected to cell transplantation under a dissecting microscope (Figure 1(Ba)) using a mouthpiece-controlled micropipette (Figure 1(Bb))
The injected solution was not spread outside the injection site (Figure 1(Bd–f)), suggesting that transplanted cells were confined within several compartments of the pancreas and may not drop out from the site
Summary
Induced pluripotent stem (iPS)/embryonic stem (ES) cells have the potential to differentiate into fully differentiated cells originating from three germ layers when they are placed under differentiation-inducing conditions [1,2]. We transplanted actively proliferating tumor cells (including iPS cells) into the pancreatic parenchyma using a mouthpiece-controlled glass micropipette under observation using a dissecting microscope to test whether these cells could grow as solid tumors in vivo. We named this new technology “intrapancreatic parenchymal cell transplantation (IPPCT)”
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