Routine heat inactivation of serum reduces its capacity to promote cell attachment

Abstract

Heat inactivation (56 degrees C for 40 min) of bovine calf serum was shown to diminish its capacity to promote the attachment of cells to plastic or glass surfaces. This effect was not observed in stationary cultures (culture dishes) but became manifest under conditions in which the cells were subjected to a small amount of liquid shear force, i.e. by growing cells in roller bottles or culture tubes. Of four cell lines tested on bovine calf serum (SV-BHK, BALB-3T3, CV-1, and FS-4) SV-BHK and CV-1 cells showed the greatest sensitivity to loss of attachment-promoting activity. Fetal bovine serum also seemed to be affected by heat inactivation but to a lesser degree than bovine calf serum. Treatment of vessel surfaces with either unheated calf serum or specific attachment factors (gelatin, poly-D-lysine, and fibronectin) greatly increased cell attachment in the presence of heat inactivated serum. Heat inactivation did not seem to affect the ability of cells to grow after attachment. Of the four cell lines tested, the normal human fibroblast line (FS-4) was shown to be most effective at conditioning medium and restoring its capacity to promote the attachment of all four cell lines.

Effects of serum and serum heat-inactivation on human bone derived osteoblast progenitor cells

“…Latter layer also plays an important role in in vitro studies. It has been noted that as result of serum heat inactivation, the cell±substratum surface interaction is changed [23]. Our present and previous study [2] indicate that HBMC performance and by that the functional state of the cells is changed by using FCS, FCSH or HSH instead of untreated HS.…”

Enumeration of the colony-forming units–fibroblast from mouse and human bone marrow in normal and pathological conditions

“…Heating serum, as a means of complement inactivation and prevention of complement-mediated cell lysis, has been regarded as a mandatory step for a wide range of cell cultures (Giard, 1987; Leshem et al, 1999). Yet FBS heat inactivation was found to be unnecessary for immune responses in vitro , such as lymphocyte proliferation, IL-2 production, and cell-mediated cytotoxicity (Leshem et al, 1999), as well as for cell attachment to plastic (Giard, 1987). Beyond such limited data, the effect of FBS heat inactivation on cell behavior has long been neglected (Mannello and Tonti, 2007).…”

An alkaline follicular fluid fraction induces capacitation and limited release of oviduct epithelium-bound stallion sperm

“…Freshly filtered (2 mm diameter) FF (pHz7.2) was treated in five different ways: i) centrifuged through a 100 kDa filter (10 min at 14 000 g; Amicon Ultra-0.5 ml Centrifugal Filters for Protein Purification and Concentration: Millipore) to see whether the effects of whole FF were elicited by molecules !100 kDa; ii) by heat inactivation in a 55 8C water bath for 0.5 h (Triglia & Linscott 1980, Giard 1987, Pinyopummintr & Bavister 1994 to examine whether the FF factor was heat-resistant; iii) charcoal treatment as described by Cheng et al (1998) to examine whether a lipid or lipid-bound factor, including steroid hormones (Quirk & Fortune 1986), was implicated. For charcoal treatment, pooled FF was stirred at ambient temperature for 45 min with 50 mg charcoal/ml (Norit: activated and neutralized; Sigma) and then centrifuged at 4500 g for 1 h at 4 8C.…”