The Utilization of Animal Product-Free Media and Autologous Serum in an Autologous Dermal Substitute Culture

Morimoto, Naoki; Takemoto, Shuhei; Kanda, Norikazu; Ayvazyan, Artem; Taira, Masaki; Suzuki, Shigehiko

doi:10.1016/j.jss.2009.11.724

Cited by 19 publications

(14 citation statements)

References 27 publications

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“…*0.01 \ p \ 0.05, **p \ 0.01; no asterisk means no significant difference (after applying for Bonferroni correction). Error bar means SD (N = 9) Cytotechnology (2015) 67:507-514 511 to Eagle's minimal essential medium: HEPES, L-glutamine, lipids, sodium bicarbonate, sodium selenite, gentamicin sulfate, non-essential amino acids, recombinant human insulin, recombinant human epidermal growth factor, dexamethasone, and synthetic polymer (Itoh and Hoshi 2005;Yamada et al 2006;Morimoto et al 2011;Takami et al 2008). It contains no animal-derived components and is thought to promote a normal fibroblast phenotype.…”

Section: Discussionmentioning

confidence: 99%

“…Currently, Dulbecco's modified Eagle medium (DMEM) plus fetal bovine serum (FBS) is usually used for human fibroblast culture (Rittié and Fisher 2005). An autologous serum can be used in place of FBS (Itoh and Hoshi 2005;Yamada et al 2006;Nishi et al 2010;Morimoto et al 2011). Fibroblasts cultured with serum have a high proliferative capacity, similar to that of keloids; however, human fibroblasts in living dermis rarely proliferate and maintain homeostasis of the dermis through collagen production.…”

Section: Introductionmentioning

confidence: 99%

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Use of synthetic serum-free medium for culture of human dermal fibroblasts to establish an experimental system similar to living dermis

et al. 2014

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In this study, we sought to establish a defined experimental system for fibroblast growth similar to that of the living dermis. To this end, we evaluated the growth and biochemical characteristics of fibroblasts cultured with serum-free HFDM-1, a finely tuned synthetic medium for human fibroblast culture. Three culture conditions were used to grow fibroblasts obtained from primary culture: (1) culture with Dulbecco's modified Eagle medium (DMEM) plus 10 % fetal bovine serum (serum-supplemented DMEM), (2) culture with DMEM (serum-free DMEM), and (3) culture with HFDM-1 (HFDM-1), and fibroblast morphology, growth, collagen type I production, and lipid composition were analyzed. Fibroblasts grown in HFDM-1 maintained cell numbers at nearly 100 % from days 14 to 21 and produced more collagen type I than cells grown in serum-supplemented and serum-free DMEM. Arachidonic acid (20:4) and total polyunsaturated fatty acids were lower in cells grown in serum-free DMEM and HFDM-1 than in serum-supplemented DMEM. These results suggested that HFDM-1 recapitulated growth conditions in the dermis better than traditional, serum-supplemented DMEM. In addition, the controlled chemical composition of HFDM-1 eliminated a potential source of variability in cell culture conditions.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Use of synthetic serum-free medium for culture of human dermal fibroblasts to establish an experimental system similar to living dermis

et al. 2014

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“…They demonstrated increasing both cell quantity, purity and reducing probable common risks. At the present study, we used human autologous serum instead of FBS similar to some of previous studies (Kamil et al 2007;Mazlyzam et al 2008;Morimoto et al 2010). There are some types of pharmagrade FBS in the country that are safer than others, but according to previous description in clinical grade procedures, the utilization of xeno-free media is the first priority of the cell manufacturer.…”

Section: Discussionmentioning

confidence: 99%

“…FDA approved some products with animal components have been safely used in humans, however, long-term hypothetical rejection and immunological reaction issues could be minimized and/or eliminated by the use of animal-free serum (Kamil et al 2007). Therefore, it is expected of culture cells which are intended to cell therapy under specific conditions and utilization of xeno-free media and serum such as human serum for achieving more safety (Lopez and De Vries 1999;Mizuno et al 2006;Mazlyzam et al 2008;Pytlik et al 2009;Morimoto et al 2010). The most important difference between this study and others is that we tried to do it as a clinical grade Schwann cell cultivation under a clean condition and avoiding the use of FBS, growth factors and cell passaging to achieve high level biosafety standards.…”

Section: Introductionmentioning

confidence: 99%

Clinical grade cultivation of human Schwann cell, by the using of human autologous serum instead of fetal bovine serum and without growth factors

et al. 2011

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Clinical grade cultivation of human schwann cell by the utilization of human autologous serum instead of fetal bovine serum, and also avoiding any growth factors, can increase safety level of this procedure in cases of clinical cell transplantation. The aim of this study was demonstration of the feasibility of clinical grade schwann cell cultivation. In this experimental study after obtaining consent from close relatives we harvested 10 sural nerves from brain death donors and then cultured in 10 seperated culture media plus autologous serum. We also prepared autologous serum from donor's whole blood. Then cultured cells were evaluated by S100 antibody staining for both morphology and purity. Cell purity range was from 97% to 99% (mean=98.11 ± 0.782%). Mean of the cell count was 14,055.56 ± 2,480.479 per micro liter. There was not significant correlation between cell purity and either the culture period or the age of donors (P>0.05). The spearman correlation coefficient for the cell purity with the period or the age of donors was 0.21 and 0.09, respectively. We demonstrated the feasibility of clinical grade schwann cell cultivation by the using of human autologous serum instead of fetal bovine serum and also without the using of growth factors. We also recommended all cell preparation facilities to adhere to the GMP and other similar quality disciplines especially in the preparation of clinically-used cell products.

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“…[42][43][44] Compare to PRP technology, isolating autologous GFs by PRF technology is considered simpler; it also contains various GF types which required for stimulation of healing processes as shown in our experiment. Similar result has been reported by Jørgensen et al, 45 in their pilot study where PRF has also effective for recalcitrant chronic wound.…”

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confidence: 95%

The Effect of PRF on Serum Starved Human Dermal Fibroblast

2016

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Healing failure on chronic ulcers was suspected due to the decrease of Growth Factors (GFs) supply caused by either GFs trapped in the fibrin, or degraded by protease, or decreased level due to reduction of GFs gene expression. Administration of various GFs can stimulate healing of chronic ulcers. High level of GFs is available in biologic material called Platelet Rich Fibrin (PRF). This study was conducted to have in vitro evidence of PRF effect on GFs-serum starved human dermal fibroblasts as representative cells of chronic ulcers. Human dermal fibroblasts (HDFs) were isolated from foreskin of six boys aged 11-14 years-old. After 24 hours of serum deprivation, HDFs were treated by 100, 50, and 25% PRF lysate diluted in cultured medium. Cellular migration was measured using scratch assay, while cellular viability was measured using MTT assay and collagen deposition was measured using Sirius Red assay. The HDFs of serum starvation group showed significant impairment activities in terms of cellular migration (25%), cellular proliferation (20%), and collagen deposition (10%) (p<0.05). Administration with various levels of PRF lysate could significantly recover those activities (p < 0.05). Because cellular activities of serum starved HDFs is similar with fibroblasts isolated from the bottom of chronic ulcers and administration of various levels of PRF lysate was capable to recover those activities, it can be concluded that PRF is a good biologic material to stimulate healing of chronic ulcers. However, in order to get better evidence based medicine, both pre clinical and clinical studies must be performed. ABSTRAKKegagalan sembuh pada ulkus khronis disebabkan oleh rendahnya suplai growth factor (GF) akibat GF terjebak dalam fibrin, atau dirusak oleh protease bakteri, atau akibat menurunnya ekspresi gena penyandi GF. Pemberian GF dapat memacu penyembuhan ulkus khronis. Berbagai GF ternyata terdapat dalam material biologis seperti fibrin kaya platelet (FKP). Penelitian ini dilakukan untuk memperoleh bukti in vitro bahwa FKP dapat mengembalikan fungsi fibroblast tanpa serum sebagai model fibroblas dari ulkus khronis. Fibroblas dermis manusia (FDM) diisolasi dari kulit kulup 6 anak laki-laki berumur 11-14 tahun. Setelah dibebaskan dari serum selama 24 jam, FDM kemudian diberi 100, 50, 25% lisat FKP dan medium pelarut sebagai kontrol. Selanjutnya, aktivitas selular berupa timbunan kolagen, proliferasi dan migrasi sel diukur berturut-turut dengan metode Sirius Merah, MTT, dan goresan. FDM tanpa serum ternyata secara mengalami penurunan kemampuan menimbun kolagen sebesar 10%, proliferasi sebesar 20% dan penurunan kemampuan migrasi sebesar 25% secara signifikan (p<0,05). Penambahan lisat FKP 111Radiono, The effect of platelet rich fibrin (PRF) on serum starved human dermal fibroblast ternyata dapat memulihkan aktivitas selular FDM tersebut. Dari eksperimen ini dapat disimpulkan bahwa FKP merupakan material biologis yang dapat dikembangkan untuk memacu penyembuhan ulkus khronis. Namun demikian, untuk memperoleh bukt...

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The Utilization of Animal Product-Free Media and Autologous Serum in an Autologous Dermal Substitute Culture

Cited by 19 publications

References 27 publications

Use of synthetic serum-free medium for culture of human dermal fibroblasts to establish an experimental system similar to living dermis

Use of synthetic serum-free medium for culture of human dermal fibroblasts to establish an experimental system similar to living dermis

Clinical grade cultivation of human Schwann cell, by the using of human autologous serum instead of fetal bovine serum and without growth factors

The Effect of PRF on Serum Starved Human Dermal Fibroblast

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