Collection Protocol of Fetal and Foreskin-derived Fibroblasts

Embryonic fibroblasts, the major cellular component of sparse connective tissue, are differentiated from mesenchymal cells during the embryonic period.
Fibroblasts are large, well-defined, mostly spindle-shaped or star-shaped flat structures with regular ovoid nuclei and large and obvious nucleoli. Fibroblasts have high functional activity, weak basophilic cytoplasm, and obvious protein synthesis and secretion activities.

Take about 3 cm² of aborted fetal abdominal skin or foreskin, place it in a 35 mm Petri dish containing DPBS, and cut it into 1 mm².
The tissue pieces are collected and transferred to a 15 mL centrifuge tube, and centrifuged at 1000 r/min for 3 min at room temperature.
Discard the supernatant, resuspend the tissue blocks with cell base culture medium, and transfer to a T25 culture flask, trying to evenly distribute the tissue blocks, while carefully aspirating the culture medium around the tissue blocks.
Invert the culture flask overnight, turn it over the next day, and add 1.5 mL of cell base culture medium.
The culture medium is replaced with a fresh culture medium every other day, and cells are seen crawling out around the tissue block for about 7 days.
Passage with 0.25% trypsin.

Foreskin tissue is taken and rinsed with DPBS containing 2% P/S.
The foreskin tissues are well cut with ophthalmic scissors, added with 0.25% trypsin, and digested until the tissues are in a mucous flocculent state.
The digestion is terminated by adding an equal volume of fibroblast culture medium (DMEM medium, 10% FBS, 1% NEAA, 1% P/S).
The cells are blown and mixed, centrifuged and resuspended, then inoculated in culture flasks and incubated in the incubator.
1-3 generations of foreskin fibroblasts are selected as the donor cells for iPSCs.

The skin and foreskin of the aborted fetus are taken from the hospital. Signed informed consent is required.

For research use only. Not for any other purpose.