Company: FTII
Filing Date: 2025-02-14
Form Type: S-4
Source: 0001493152-25-006997
Chunk: 346

Company: FutureTech II Acquisition Corp.
Filing Date: 2025-02-14
Form: S-4
Chunk 346
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 clinical trial. Npj Regenerative Medicine, 7(1). https://doi.org/10.1038/s41536-021-00197-1 |

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We evaluated the chemo-attractive
properties of AAT in vitro studies by measuring the migration of human adipose derived stem cells (“ASCs”) through a
porous membrane in response to soluble molecular signals. Serum-starved human ASCs were screened in a Boyden chamber for 6 hours to
determine whether they would migrate towards adipose ECM proteins. Serum-free media was used as the negative control for cell
migration and media supplemented with 10% fetal bovine serum (“FBS”) was used as the positive control. A significant
increase in the number of cells migrating across the membrane was observed with the addition of 1% AAT to serum-free media in the
lower chamber, or 51.8% of the migration observed with the positive control of 10% FBS. In addition to attracting ASCs with soluble
factors, AAT provides a physical substrate for cell attachment and differentiation. ASCs seeded on AAT-coated slides preferentially
adhered to the matrix proteins over areas of exposed glass and adapted a spindle-shaped mesenchymal morphology.

With the addition of adipogenic induction
media, cells underwent adipogenesis as determined by the adoption of a round morphology characteristic of mature adipocytes and accumulation
of lipid droplets that stained positively with Nile Red. To determine if the AAT scaffold provided a unique substrate for adipogenesis,
we compared the adipogenic differentiation potential of ASCs cultured in AAT and acellular dermis (“ACD”) in a 3D culture
environment. Constructs were initially maintained in growth media for 3 days after which they were transferred into adipo-inductive media
until day 15. After 15 days, AAT and ACD constructs looked indistinguishable upon gross examination. However, histological staining with
Oil Red O revealed greater lipid accumulation in the AAT construct compared to ACD. Furthermore, gene expression of early (PPARG and CEPBA)
and late (FABP4, LPL, LEP) markers of adipogenesis in ASCs significantly increased in the AAT compared to ACD.

To test different
batches of human-derived AAT for in vivobiocompatibility, subcutaneous injections were carried out in Sprague Dawley rats (n =
12) for up to twelve weeks. Animals were grouped