BackgroundIn-vitro spermatogenesis in mammalian species is considered an important topic in reproductive biology. New strategies for achieving a complete version of spermatogenesis ex vivo have been conducted using an organ culture method or culture of testicular cells in a three-dimensional soft agar culture system (SACS). The aim of this study was to develop a new method that supports spermatogenesis to the meiotic phase and morphologically mature spermatozoa through the culture of testicular cells and seminiferous tubules (STs) in a modified SACS, respectively.MethodsFirst, enzymatically dissociated testicular cells and mechanically dissociated STs of neonatal mice were separately embedded in agarose and then placed on the flat surface of agarose gel half-soaked in the medium to continue culture with a gas-liquid interphase method.ResultsFollowing 40 days of culture, the meiotic (Scp3) and post-meiotic (Acr) gene expression in aggregates and STs was confirmed by real-time polymerase chain reaction. These results were complemented by immunohistochemistry. The presence of morphologically mature spermatozoa in the frozen sections of STs was demonstrated with hematoxylin and eosin staining. We observed Plzf- or Integrin α6-positive spermatogonia in both cultures after 40 days, indicating the potency of the culture system for both self-renewal and differentiation.ConclusionsThis technique can be used as a valuable approach for performing research on spermatogenesis and translating it into the human clinical setting.
Continual improvement of product quality has been a long challenge to Semi-Solid Metal
(SSM) technology. By conventional semi-solid processes, this might be attained at the expense of
economical production. The advent of Inclined Cooling Plate (ICP) process has already realized the
development of non-dendritic SSM while satisfying qualitative, quantitative and economical
requirements collectively. In spite of its potential advantages, functional mechanisms of this process
are not yet clearly understood that makes its optimal utilization obscured.
Basically, such understanding needs a picture of the process. As the first step, this picture is pursued
through physical modeling of the ICP process i.e. direct observation of an analog system by virtue
of transparent character of a model alloy (succinonitrile-acetone). Based on this phenomenological
model, a picture of the process is presented as follows: flowing molten alloy down ICP, multiple
regions form typically on the plate i.e. a chilled layer at the vicinity of the plate surface, a two-phase
mushy zone on the chilled layer and ambient liquid far from the plate surface. In this process,
interaction of the liquid forced-flow with mushy zone separates solid particles from the stationary
mush on the plate resulting in a two-phase mixture which is responsible for the formation of slurry
i.e. SSM.
Semi-solid forming (SSF) involves alloys with non-dendrite microstructure that contain spherical
solid particles in the liquid matrix. This process is generally divided into three main steps: feedstock
manufacturing, reheating and forming.
Feedstock has the main effect on the cost and quality of product. Many researches have been carried
out to reduce the cost of feedstock manufacturing. Slope plate is a simple semi-solid process that
can reduce the cost of feedstock. In this study a Cu plate with water and without water circulation
was used to investigate the effect of pouring temperature, cooling rate during casting and
solidification in the mold on the microstructure. Cast ingots with optimized microstructure were
reheated in various time and temperatures in semi-solid region to obtain kinetics of globularization
and solid grain growth.
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