234: Growth factors in amniotic fluid remain bioavailable after storage

Necrotizing enterocolitis (NEC) is the most common gastrointestinal (GIT) complication of prematurity. In utero, the fetus swallow amniotic fluid (AF) which has growth factors (GFs). These GFs are responsible for maturation & development of GIT. In vitro and animal studies have shown that GFs in AF have trophic effect on GIT epithelium & are protective against NEC. Whether the GFs in AF are biologically active ex utero, remains to be seen. In order to study this, we need to determine if GFs in AF remain active after storage ex-utero for at least 7 days. We did a prospective study to find out if AF can be stored for 7 days in fridge without deterioration of activity of different GFs. Preterm infants are generally too sick to be breast fed breast milk (good source of GFs). But AF can be given to premature infants because in-utero as fetus they were swallowing AF. Hence AF can be alternative good source of GFs for first week of life when they are prone to develop NEC. We collected AF from term pregnant women undergoing elective cesarean section. After collection of AF, 10 mL was stored in fridge for seven days (post storage sample) and 10 mL was stored at −80 degree. Human fetal small intestinal cells (FHs 74) were exposed to different concentrations of AF (10, 20, 30, 40%) and proliferation of cells was assessed by WST-1 assay. Then FHs 74 cells were exposed to pre & post storage amniotic fluid to show the biological activity of GFs after storage. We found that FHs-74 cell proliferation was maximum when they were exposed to 30% of AF & proliferation was significantly higher as compare to no AF. When FHs 74 cells were exposed to pre & post storage AF (30%) there was no difference in the cell proliferation assessed by WST-1 assay. Human AF can be stored in fridge for at least for one week without deterioration of the biological activity of GFs present in AF. Human AF may be a good source of GFs for extremely premature infants during first week to decrease and/or prevent NEC. Further studies may be required before using AF.