Chickpea protein hydrolysate as a substitute for serum in cell culture

Abstract

The growth of mammalian cells in vitro requires the use of rich culture media that are prepared by combining serum with specific nutrient formulations. Serum, the most expensive component of culture media, provides a complex mixture of growth factors and nutrients. Protein hydrolysates that can support in vitro cell growth and eliminate or reduce the need to use serum have been obtained from different sources. Here we describe the use of two food grade proteases to produce a chickpea protein hydrolysate that has been added to cell culture medium in order to determine whether it can be used as a substitute for serum. Medium containing the hydrolysate has been tested using two human cells lines: the monocytic THP-1 cell line which grows in suspension, and the epithelial Caco-2 cell line which grows as a monolayer. The chickpea protein hydrolysate was a good substitute for serum in the first case, but did not allow growth of Caco-2 cells. Supplementation of culture media with this inexpensive and safe hydrolysate would greatly reduce the cost of cell culture.

Effect of chickpea protein hydrolysates on cell proliferation and in vitro bioavailability

“…Thus, THP-1 cells kept growing after 20 passages when cultured in medium supplemented with alcalase-flavourzyme hydrolysate in the absence of serum, but Caco-2 cells did not grow at all (Giron-Calle et al, 2008). Nevertheless, no correlation could be found between the long-term and short-term effects on growth in the absence of serum.…”

“…Thus, proliferation of THP-1 cells was actually inhibited by up to 100% in cultures exposed to hydrolysates with various DH for up to 3 days. In the presence of serum, proliferation of Caco-2 cells was inhibited by up to 30% under certain conditions, while proliferation of THP-1 cells was actually promoted by up to 100% in the presence of 0.5% FBS, and no effect on proliferation was observed when the THP-1 cells were exposed to hydrolysates in the presence of 10% FBS (Giron-Calle et al, 2008). Comparing these results with the present study, it should be concluded that although enzymatic hydrolysis of chickpea protein using either microbial or physiological enzymes produces growth-promoting and growth-inhibiting peptides, the activity profile of hydrolysates with the same DH produced with either type of enzymes is not the same.…”

“…Isolates were further washed with acetone, ethyl ether, ethanol, and water, twice each, by resuspending at 20% (v/v) in these solvents and discarding supernatants after centrifugation at 1000g and 22°C. The resulting isolates were 98% (w/w) protein (Giron-Calle et al, 2008;Girón-Calle et al, 2004).…”

“…It has previously been described that chickpea hydrolysates obtained by treatment with two food-grade microbial protease preparations, alcalase and flavourzyme (Novozymes, Bagsvaerd, Denmark), can substitute for serum in cell culture and sustain growth of THP-1 but not Caco-2 cells, although the effects on short-term growth were mixed (Giron-Calle et al, 2008). Cell proliferation under exposure to chickpea protein hydrolysates that were produced by treatment with the physiological proteases pepsin and pancreatin has now been studied using the same cells lines.…”

Food‐derived immunomodulatory peptides

“…A related study reported on the production of hydrolysates of chickpeas by microbial proteases; the obtained peptides promoted the proliferation of human monocyte THP-1 cells but inhibited the proliferation of Caco-2 cells. 130 Subsequently, the authors evaluated the effect of hydrolysates obtained with pepsin and pancreatin on the proliferation of THP-1 and Caco-2 cells. The hydrolysates inhibited the proliferation of Caco-2 cells up to 45% and of THP-1 cells up to 78%.…”