To evaluate the role of growth factors in amniotic fluid and in human milk on gastrointestinal adaptation of the fetus and very low-birth-weight infants, the effects of these fluids and multiple growth factors were investigated in a human fetal small intestinal cell line (FHs 74 Int).
After FHs 74 Int cells were incubated with amniotic fluid, human milk, or recombinant growth factors, growth-promoting activity was measured by [3H]-thymidine incorporation into cells.
Incubating cells with amniotic fluid or human milk promoted growth dose dependently. Genistein almost completely inhibited growth-promoting activity in amniotic fluid P = 0.002), and growth was partially inhibited by antibodies against epidermal growth factor (EGF) (P = 0.047), insulin-like growth factor-1 (IGF-1, P = 0.047), or fibroblast growth factor (FGF, P = 0.014). This activity in human milk was inhibited almost completely by genistein (P < 0.0001) and partially inhibited by antibodies against EGF (P = 0.036), IGF-1 (P = 0.009), FGF (P = 0.004), hepatocyte growth factor (HGF, P = 0.001), or transforming growth factor-alpha (TGF-alpha, P = 0.001). Although recombinant EGF, IGF-1, FGF, HGF, and TGF-alpha elicited a synergistic trophic response on cultured cells, the response was much less than with amniotic fluid or with human milk.
In aminiotic fluid and in human milk, EGF, IGF-1, FGF, HGF, and TGF-alpha have a strong trophic effect on immature intestinal cells and may be involved in perinatal gastrointestinal adaptation.
"Because not all mothers of premature infants produce sufficient milk to supply their infants throughout hospitalization, it remains an important objective to define the nature and amount of enteral diet that is required postnatally to facilitate optimal intestinal tolerance. Similar to breast milk, amniotic fluid may contain important immunomodulatory, growth and antimicrobial factors (Akinbi et al., 2004; Hirai et al., 2002) that may act similarly to facilitate tolerance to bacterial antigens and dampen excessive TLR-mediated responses in the perinatal period. Continuation of fetal amniotic fluid intake after birth, as a mode of minimal enteral nutrition during TPN, has been suggested (Barney et al., 2007), but never tested in controlled studies, partly due to obvious practical difficulties in procuring un-interrupted supplies. "
[Show abstract][Hide abstract] ABSTRACT: Necrotizing enterocolitis (NEC) is the most common gastrointestinal complication in human neonates, yet the pathogenesis of this disease remains poorly understood. A fundamental approach to understanding the etiology and underlying biology of NEC is the use of in vivo experimental animal models, primarly neonatal rodents and pigs. The rodent models using rats and mice have provided a much of the experimental evidence showing the protective influence of breast milk and the role of specific molecular mechanisms involved in the premature innate immune and intestinal injury response. A key advantage of mice is the abilty to test how genetic disruption of specific genes alters the NEC phenotype. More recently, pigs have emerged as an animal model of NEC and used to establish the role of bacterial colonization, prematurity, parenteral nutrition and antibiotic therapy. This review will outline some of the advantages and disadvantages of both rodent and pig models and highlight the lessons learned about NEC pathobiology from these different experimental models.
"Compared to other growth factors, HAF is less expensive and more readily available. This fluid is a natural and enriched human source of various growth factors with synergistic effects (Underwood et al., 2005; Hirai et al., 2002; Park et al., 2006). HAF has an approximately neutral pH (7.2), and its osmotic pressure is in the physiological range. "
[Show abstract][Hide abstract] ABSTRACT: The present study was designed to evaluate the effects of human amniotic fluid (HAF) on the growth of human corneal endothelial cells (HCECs) and to establish an in vitro method for expanding HCECs. HCECs were cultured in DMEM-F12 supplemented with 20% fetal bovine serum (FBS). Confluent monolayer cultures were trypsinized and passaged using either FBS- or HAF-containing media. Cell proliferation and cell death ELISA assays were performed to determine the effect of HAF on cell growth and viability. The identity of the cells cultured in 20% HAF was determined using immunocytochemistry (ICC) and real-time reverse transcription polymerase chain reaction (RT-PCR) techniques to evaluate the expression of factors that are characteristic of HCECs, including Ki-67, Vimentin, Na+/K+-ATPase and ZO-1. HCEC primary cultures were successfully established using 20% HAF-containing medium, and these cultures demonstrated rapid cell proliferation according to the cell proliferation and death ELISA assay results. The ICC and real time RT-PCR results indicated that there was a higher expression of Na+/K+-ATPase and ZO-1 in the 20% HAF cell cultures compared with the control (20% FBS) (P<0.05). The 20% HAF-containing medium exhibited a greater stimulatory effect on HCEC growth and could represent a potential enriched supplement for HCEC regeneration studies.
Experimental Eye Research 04/2014; 122. DOI:10.1016/j.exer.2014.04.002 · 2.71 Impact Factor
"From the 14 th to 16 th weeks of gestation, HAF consists of multiple trophic factors essential for fetal growth and embryonic cell proliferation and differentiation  . For example, Hirai et al. (2002) found that multiple growth factors in HAF promote the growth of human fetal small intestinal cells in culture . It also seems that some HAF components, such as fibronectin (the 3 rd most abundant protein in HAF in the 16 th week of gestation ), plays a pivotal role in cell attachment, polarity, and migration . "
[Show abstract][Hide abstract] ABSTRACT: Retinal pigment epithelial (RPE) cells are capable of differentiating into retinal neurons when induced by the appropriate growth factors. Amniotic fluid contains a variety of growth factors that are crucial for the development of a fetus. In this study, the effects of human amniotic fluid (HAF) on primary RPE cell cultures were evaluated.
RPE cells were isolated from the globes of postnatal human cadavers. The isolated cells were plated and grown in DMEM/F12 with 10% fetal bovine serum. To confirm the RPE identity of the cultured cells, they were immunocytochemically examined for the presence of the RPE cell-specific marker RPE65. RPE cultures obtained from passages 2-7 were treated with HAF and examined morphologically for 1 month. To determine whether retinal neurons or progenitors developed in the treated cultures, specific markers for bipolar (protein kinase C isomer α, PKCα), amacrine (cellular retinoic acid-binding protein I, CRABPI), and neural progenitor (NESTIN) cells were sought, and the amount of mRNA was quantified using real-time PCR.
Treating RPE cells with HAF led to a significant decrease in the number of RPE65-positive cells, while PKCα- and CRABPI-positive cells were detected in the cultures. Compared with the fetal bovine serum-treated cultures, the levels of mRNAs quantitatively increased by 2-, 20- and 22-fold for NESTIN, PKCα, and CRABPI, respectively. The RPE cultures treated with HAF established spheres containing both pigmented and nonpigmented cells, which expressed neural progenitor markers such as NESTIN.
This study showed that HAF can induce RPE cells to transdifferentiate into retinal neurons and progenitor cells, and that it provides a potential source for cell-based therapies to treat retinal diseases.
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