Method for evaluating quality of cultured neonatal pig Sertoli cells

Abstract

We describe an alternative strategy for preparing and identifying cultured SC for further assays of metabolic activity or in transplantation models. Establishing a one-step Liberase-digestion method for isolation, evaluating viability and apoptosis by more sensitive methods, and detecting specific markers in culture can help to evaluate the quality of cultured cells. Specific cell markers for identifying SC may be critical when identifying SC outside the testis, in contrast with vimentin which is useful only for in situ cells.

Reversal of experimental Laron Syndrome by xenotransplantation of microencapsulated porcine Sertoli cells

“…To detect the presence of Müllerian inhibiting substance (MIS), a unique pre-pubertal SC marker, anti-MIS immunostaining was performed according to previously reported methods with minor changes [10,21,22]. Briefly, pSC monolayers were fixed in 4% paraformaldehyde-phosphatebuffered saline (PBS) for 30 min.…”

Correlation between insulin requirements and anti-galactose antibodies in patients with type 1 diabetes transplanted with neonatal pig islets

“…The devices consisted of 6 × 0·8‐cm surgical‐grade stainless steel mesh tubes, with a polytetrafluoroethylene (PTFE) rod inside. Two months after implanting the devices, the transplant procedure was carried out by exposing the end of each device, removing the PTFE rod and infusing 250 000 islets of Langerhans and 30–100 Sertoli cells per islet into the collagen tube with improved methods of Sertoli cells and neonatal islet porcine cultures [20,21]. Each patient received a mean of three cellular infusions, with a total of 68.…”

Xenograft of Microencapsulated Sertoli Cells Reverses T1DM in NOD Mice by Inducing Neogenesis of Beta-Cells

“…Mullerian inhibiting substance immunostaining was performed according to previously established methods (21,22). Endogenous alkaline phosphatase activity was assessed, according to Chapin's procedure, to detect peritubular cells (23).…”