Supplementation of GelMA with Minimally Processed Tissue Promotes the Formation of Densely Packed Skeletal-Muscle-Like Tissues.

We present a simple and cost-effective strategy for developing gelatin methacryloyl (GelMA) hydrogels supplemented with minimally processed tissue (MPT) to fabricate densely packed skeletal-muscle-like tissues. MPT powder was prepared from skeletal muscle by freeze-drying, grinding, and sieving. Cell-culture experiments showed that the incorporation of 0.5-2.0% (w/v) MPT into GelMA hydrogels enhances the proliferation of murine myoblasts (C2C12 cells) compared to proliferation in pristine GelMA hydrogels and GelMA supplemented with decellularized skeletal-muscle tissues (DCTs). MPT-supplemented constructs also preserved their three-dimensional (3D) integrity for 28 days. By contrast, analogous pristine GelMA constructs only maintained their structure for 14 days or less. C2C12 cells embedded in MPT-supplemented constructs exhibited a higher degree of cell alignment and reached a significantly higher density than cells loaded in pristine GelMA constructs. Our results suggest that the addition of MPT incorporates a rich source of biochemical and topological cues, such as growth factors, glycosaminoglycans (GAGs), and structurally preserved proteins (e.g., collagen). In addition, GelMA supplemented with MPT showed suitable rheological properties for use as bioinks for extrusion bioprinting. We envision that this simple and cost-effective strategy of hydrogel supplementation will evolve into an exciting spectrum of applications for tissue engineers, primarily in the biofabrication of relevant microtissues for in vitro models and cultured meat and ultimately for the biofabrication of transplant materials using autologous MPT.

Estrus induction in anestrous mixed-breed goats using the "female-to-female effect".

A trial was conducted during the anestrous period in female goats to determine: (a) whether estrus can be induced in anestrous goats by administration of equine chorionic gonadotropic hormone (eCG) and PGF2α under pen conditions and (b) whether these sexually active female goats can elicit sexual arousal in sexually inactive bucks. One hundred and fifteen pluriparous, nonlactating mixed-breed female goats were randomly assigned to one of four treatment groups: (1) administration of a single dose of 240 IU of eCG, 50 µg PGF2α i.m., and 25 mg progesterone (P4) (eCG; n=30); (2) administration of P4 and exposure to female goats treated with eCG-PGF2α (P4; n=39); (3) administration of 0.5 ml saline and P4 (Sal; n=23); and (4) P4 plus exposure to female goats treated with saline (Con; n=23). After hormone administration, all goats were put together with adult sexually inactive bucks for 15 days. The percentage of goats in estrus during these 15 days was similar in eCG-treated animals and untreated animals exposed to the eCG animals (97 and 95%). Pregnancy rate was also similar (63 vs. 64%) between these two groups. eCG-treated goats exhibited estrus earlier (P<0.05) than the treated goats in contact with the eCG goats. Furthermore, eCG-treated goats had larger litters (1.9 ± 0.2 vs. 1.6 ± 0.1, P<0.05) than the untreated goats in contact with the eCG goats. These results show that fertile estrus can be induced in anestrous female goats by exposing them to female goats induced to estrus with eCG. This female-female interaction triggers the stimulation cycle leading to the sexual arousal of bucks.