산처리 공정에 따라 추출한 돈피 젤라틴의 이화학적 특성에 관한 연구

염건웅,민상기,Yeom Geun-Woong,J Andrieu,Min Sang-Gi 한국축산식품학회 2004 한국축산식품학회지 Vol.24 No.3

The objective of this study was to investigate the physicochemical characteristics of gelatin extracted from pork skin under soaking in various acid solutions (lactic acid, acetic acid, and citric acid). Gelatin sol was extracted at 8$0^{\circ}C$, frozen at -2$0^{\circ}C$ and lyophilized it for 3 days to be completely dried in freeze drying unit. In the evaluation of gelatin quality, gelatin soaked in citric acid showed higher L- and a-values than those of any other gelatin (p<0.05). Gelatin treated by acetic acid showed the highest gel strength, cohesiveness, and brittleness. The content of hydroxyproline amino acid in gelatin treated by acetic acid was larger than one of gelatin treated in lactic and citric acid in order. From the experimental results, the highest quality of gelatin in all of period, which was soaked in acetic acid and lactic acid, has a more good quality than gelatin soaked in citric acid. 본 연구는 gelatin의 생산공정에서 각종 산 처리가 최종 gelatin의 품질에 어떤 영향을 주는지를 평가함으로써 양질의 gelatin을 생산하는데 기초를 제공하고자 하였다. Gelatin gel의 물성은 lactic acid로 처리한 gelatin이 높은 hardness, cohesiveness, brittleness를 나타내었고 다음으로는 citric acid, acetic acid 순으로 나타났다. Gelatin의 색도는 citric acid, 처리한 gelatin이 acetic acid나 lactic acid로 산침한 젤라틴 보다 높은 명도를 나타냈다. a-value는 acetic acid로 산침한 gelatin이 가장 높게 나타났으며 황색도를 나타내는 b-value는 lactic > acetic acid > citric acid 순으로 높게 나타났다. Gelatin의 점도를 측정한 결과 전단속도(shear rate)에 대한 전단응력(shear stress)의 관계는 모두 전단속도가 증가함에 따라 전단응력이 증가하는Newton성 유체로 판단되었다. 그 기울기는 acetic acid로 산침한 것이 가장 높게 나타났으며 lactic acid, citric acid 순으로 높게 나타났다. Gelatin내의 hydroxyproline 함량을 살펴보면 acetic acid(0.186 g/100 g sample)로 가장 높은 수치를 나타나며 다음으로 lactic acid, citric acid 순으로 나타났다. 그러나, sample중 crude protein 함량을 비교하면 acetic acid(68.74%), lactic acid(80.08%), citric acid(85.47%)로 나타났다. 이것은 gelatin의 열수 추출시 가장 작게 용출된 acetic acid가 가장 높은 collagenous connective tissue 함량(1.35 g/100g)을 나타내었다.

A New Method of Gelatin Modified Collagen and Viscoelastic Study of Gelatin-Collagen Composite Hydrogel

Lang He,Sheng Li,Chengzhi Xu,Benmei Wei,Juntao Zhang,Yuling Xu,Beirong Zhu,Yang Cao,Xilin Wu,Zhijin Xiong,Rongrui Huang,Jian Yang,Haibo Wang 한국고분자학회 2020 Macromolecular Research Vol.28 No.9

Pure collagen materials are expensive with poor mechanical properties, which need modifications in most cases. As the degradation product of collagen, gelatin is cheap, degradable and biocompatible, but few literatures have reported the research about gelatin-collagen composite materials. This is because gelatin and collagen have different soluble temperatures—gelatin is soluble in hot water (≥30 oC) and swells in cold water. However, a low temperature (2-10 oC) is required to prepare and store collagen solution, and neutral collagen solution denatures quickly above the room temperature. In this study, gelatin was ground into powders and swelled in neutral bovine tendon pepsin-soluble collagen solution (BPSC) to form a homogeneous gelatin-collagen mixture, in light of the swelling characteristics of gelatin in cold water. The assembly properties and gel properties of this composite material were further studied. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) test results showed that the bovine tendon collagen had typical type-I collagen structural characterizations with two α chains of about 100 kDa and one β chain of about 200 kDa; while the SDS-PAGE pattern of gelatin displayed bands continuously distributed from 30 to 200 kDa. Amino acid composition analysis test indicated that the content of polar amino acids and the sum of acidic and base amino acids for gelatin were higher than that of BPSC. Studies on gel properties demonstrated that gelatin-collagen mixed solution had collagenlike assembly characteristics and assembly kinetics. The moduli of the assembled gel at 35 oC were equivalent to that of pure bovine tendon collagen system; moreover, the system moduli didn’t change with time with elastic moduli (G') of about 40 Pa. However, at 25 oC, the moduli of gelatin-collagen composite hydrogel increased with the extension of time, its G' increased about 18 times within 8 h, and the ratio of elastic modulus to viscous modulus (G'') increased 4.6 times, showing a significant aging effect of structural strength. Meanwhile, the mechanical strength of the composite hydrogel was also regulated by temperature—the gel was highly elastic (G'≈3,000 Pa, G'>>G'') at a low temperature (5 oC); as the temperature rose, the system moduli gradually decreased and the elastic gel transformed into waterlike fluid at 50 oC little by little. What’s more, gelatin-collagen composite hydrogel also had reversible sol-gel performances and self-healing capability similar to the gelatin hydrogel. This novel preparation method for preparing composite materials and the resultant composite hydrogel are expected to be used in the fields of natural food gels, injectable hydrogels, cell scaffolds, drug sustained-release materials and so on, and improve and promote the processing performances, price and large-scale production of collagen-based materials.

Effect of Duck Feet Gelatin Concentration on Physicochemical, Textural, and Sensory Properties of Duck Meat Jellies

Kim, Hyun-Wook,Park, Jae-Hyun,Yeo, Eui-Joo,Hwang, Ko-Eun,Song, Dong-Heon,Kim, Yong-Jae,Ham, Youn-Kyung,Jeong, Tae-Jun,Choi, Yun-Sang,Kim, Cheon-Jei Korean Society for Food Science of Animal Resource 2014 한국축산식품학회지 Vol.34 No.3

This study was conducted to determine the effect of duck feet gelatin concentration on the physicochemical, textural and sensory properties of duck meat jellies. Duck feet gelatin was prepared with acidic swelling and hot water extraction. In this study, four duck meat jellies were formulated with 3, 4, 5, and 6% duck feet gelatin, respectively. In the preliminary experiment, the increase in duck feet gelatin ranged from 5 to 20%, resulting in a significant (p<0.001) increase in the color score, but a decline in the hardness and dispersibility satisfaction scores. An increase in the added amount of duck feet gelatin contributed to decreased lightness and increased protein content in duck meat jellies. Regarding the textural properties, increase in the added amount of duck feet gelatin highly correlated with the hardness in the center (p<0.01, $R^2=0.91$), and edge (p<0.01, $R^2=0.89$), of duck meat jellies. Meanwhile, the increase in duck feet gelatin decreased the score for textural satisfaction; duck meat jellies containing 6% duck feet gelatin had a significantly lower textural satisfaction score, than those containing 3% duck feet gelatin (p<0.05). Furthermore, a significant difference in the overall acceptance of duck meat jellies formulated with 5% duck feet gelatin was observed, as compared to those prepared with 3% duck feet gelatin. Therefore, this study suggested that duck feet gelatin is a useful ingredient for manufacturing cold-cut meat products. In consideration of the sensory acceptance, the optimal level of duck feet gelatin in duck meat jellies was determined to be 5%.

돈육 젤라틴의 형태와 Transglutaminase의 첨가 유무에 따른 저지방 모델 소시지의 이화학적 및 조직 특성

임경훈(Kyeong Hoon Lim),이창훈(Chang Hoon Lee),진구복(Koo Bok Chin) 한국식품영양과학회 2017 한국식품영양과학회지 Vol.46 No.8

본 연구는 지방대체재로써의 젤라틴의 형태를 달리하여 첨가하였을 때의 효과와 식품첨가제로써의 TGase의 첨가가 저지방 돈육 소시지의 품질 특성에 미치는 영향에 대해 알아보기 위해서 실시하였다. 지방대체재로써 젤라틴의 첨가는 형태에 상관없이 황색도가 높은 경향을 나타내었으나, 젤라틴을 분말 형태로 첨가한 처리구가 수화 겔 형태로 첨가한 처리구보다 수분 함량이 높은 경향을 나타내었다. 보수력에서는 수화 겔 형태로 첨가한 처리구보다 낮은 경향을 나타내었다. 따라서 젤라틴을 수화 겔 형태로 첨가한 처리구가 분말 형태로 첨가한 처리구보다 기능성 면에서 뛰어난 효과를 나타내었다. 또한, 젤라틴을 수화 겔 형태로 첨가하고 TGase를 단독 혹은 젤라틴과 복합으로 첨가하여 그 특성을 알아보았다. TGase를 단독으로 첨가한 처리구는 다른 처리구들에 비해 보수력이 낮아지는 경향을 나타내었다. 따라서 TGase와 젤라틴을 함께 첨가한 처리구는 다른 처리구들보다 경도, 검성, 씹힘성이 더 높은 경향을 나타내었다. 따라서 TGase와 젤라틴을 함께 첨가하였을 때 품질 면에서 우수한 효과를 나타내었다. The objective of this study was to evaluate the effects of gelatin type (powder vs. hydrated) with or without transglutaminase (TGase) on the physicochemical and textural properties of low-fat model sausages (LFS). Treatments included LFS (control), LFS with hydrated-gel form of gelatin (1%), and LFS with powder form of gelatin (1%). Yellowness values of LFS with any type of gelatin were higher than those without gelatin (P<0.05). Moisture content (%) of LFS containing powder form of gelatin (1%) was higher than those with hydrated-gel form of gelatin or control (P<0.05). Expressible moisture (EM, %) of LFS with hydrated-gel form of gelatin was lower than those with powder form of gelatin (P<0.05). Thus, sausages with hydrated-gel form of gelatin showed better functional properties as compared to those with powder form of gelatin. To elucidate the interaction between gelatin and TGase in meat product, five actual sausages were manufactured: reference [konjac flour (KF), carrageenan (CN), and soy protein isolate], control (KF and CN alone), TRT1 (KF and CN, TGase 1%), TRT2 (KF and CN, gelatin 1%), and TRT3 (KF and CN, TGase 1%+gelatin 1%). EM (%) of sausages with TGase alone was higher than those of other treatments (P<0.05). Most textural properties of TRT3 were higher than those of other treatments. Thus, TRT3 showed better functional properties than those with single addition. In conclusion, a combination of TGase and gelatin could be used to manufacture LFSs with improved functional and textural properties.

황다랑어 복부 껍질로부터 추출한 gelatin의 물리화학적 특성

유승재,조승목,우진욱,김상호,변상훈,김태완,김선봉 한국수산과학회 2008 한국수산과학회지 Vol.41 No.6

Physicochemical characteristics of gelatin extracted from abdominal skin of yellowfin tuna (Thunnus albacares), were investigated by comparing its proximate composition, pH, amino acid composition, viscoelastic properties, gel strength and SDS-PAGE patterns, with those of bovine and porcine gelatins. The effects of gelatin concentration, maturation time, heat and freeze treatments on the gel strength of yellowfin tuna abdominal skin gelatin were studied. Amounts of α-chains, β- and γ-components of yellowfin tuna abdominal skin gelatin were higher than those of the two mammailan gelatins. Yellowfin tuna abdominal skin gelatin had the lowest imino acids (proline and hydroxyproline) content, which was consistent with that of other fishes. However, yellowfin tuna abdominal skin gelatin was highest in glycine, alanine, and lysine. The gel strengths of all gelatins were proportional to the concentration of gelatin, but yellowfin tuna abdominal skin gelatin exhibited the greatest gel strength at each concentration. Yellowfin tuna abdominal skin gelatin required a longer maturation time than the two mammalian gelatins to form a firm gel. Higher heating temperature decreased the gel strength of yellowfin tuna abdominal skin gelatin more than in the two mammalian gelatins. Freezing decreased the gel strength of bovine gelatin only slightly, but longer freezing times resulted in greater reductions in gel strength in the yellowfin tuna abdominal skin and porcine gelatins.

Effect of Duck Feet Gelatin Concentration on Physicochemical, Textural, and Sensory Properties of Duck Meat Jellies

Hyun Wook Kim,Jae Hyun Park,Eui Joo Yeo,Ko Eun Hwang,Dong Heon Song,Yong Jae Kim,Youn Kyung Ham,Tae Jun Jeong,Yun Sang Choi 한국축산식품학회 2014 한국축산식품학회지 Vol.34 No.3

This study was conducted to determine the effect of duck feet gelatin concentration on the physicochemical, textural and sensory properties of duck meat jellies. Duck feet gelatin was prepared with acidic swelling and hot water extraction. In this study, four duck meat jellies were formulated with 3, 4, 5, and 6% duck feet gelatin, respectively. In the preliminary experiment, the increase in duck feet gelatin ranged from 5 to 20%, resulting in a significant (p<0.001) increase in the color score, but a decline in the hardness and dispersibility satisfaction scores. An increase in the added amount of duck feet gelatin contributed to decreased lightness and increased protein content in duck meat jellies. Regarding the textural properties, increase in the added amount of duck feet gelatin highly correlated with the hardness in the center (p<0.01, R2=0.91), and edge (p<0.01, R2=0.89), of duck meat jellies. Meanwhile, the increase in duck feet gelatin decreased the score for textural satisfaction; duck meat jellies containing 6% duck feet gelatin had a significantly lower textural satisfaction score, than those containing 3% duck feet gelatin (p<0.05). Furthermore, a significant difference in the overall acceptance of duck meat jellies formulated with 5% duck feet gelatin was observed, as compared to those prepared with 3% duck feet gelatin. Therefore, this study suggested that duck feet gelatin is a useful ingredient for manufacturing cold-cut meat products. In consideration of the sensory acceptance, the optimal level of duck feet gelatin in duck meat jellies was determined to be 5%.

돈피 젤라틴을 첨가한 유화형 소시지의 이화학적 특성 연구

박신영(Sin-Young Park),김계웅(Gye-Woong Kim),김학연(Hack-Youn Kim) 한국식품영양과학회 2016 한국식품영양과학회지 Vol.45 No.2

본 연구는 돈피 젤라틴 첨가에 따른 유화형 소시지의 이화학적 특성을 조사하였다. 처리구별로 유화형 소시지 제조 시 젤라틴 함량을 0%, 1%, 3%, 5%를 첨가하여 제조하였다. 일반성분 측정 결과 수분 함량은 대조구에 비해 젤라틴 3%와 5% 첨가구가 유의적으로 높았으며(P<0.05), 단백질 함량은 젤라틴 5% 첨가구가 대조구와 젤라틴 1% 첨가구보다 유의적으로 높은 결과를 나타내었다(P<0.05). 또한 지방은 감소하는 경향을 보였으나 유의적인 차이는 없었다. pH는 가열 전후 모두 젤라틴 첨가 함량이 증가할수록 유의적으로 증가하였다(P<0.05). 색도 측정 결과 가열 전 적색도와 황색도는 처리구 간에 유의적인 차이가 없었으며, 명도는 젤라틴 첨가구가 유의적으로 높은 값을 나타내었다(P<0.05). 가열 후 명도와 황색도는 젤라틴 첨가구가 대조구에 비해 유의적으로 높은 값을 나타내었으며(P<0.05), 적색도는 젤라틴 3%, 5% 첨가구가 낮은 값을 나타내었다(P<0.05). 보수력은 대조구가 젤라틴 첨가구보다 유의적으로 낮았으며(P<0.05), 젤라틴 3%와 5% 첨가구가 유의적으로 높은 값을 나타내었다(P<0.05). 가열수율은 젤라틴 첨가량이 증가할수록 유의적으로 높아졌다(P<0.05). 점도는 젤라틴 3%와 5% 첨가구가 대조구와 1% 첨가구보다 유의적으로 높았다(P<0.05). 물성 측정 결과 탄력성, 응집성, 씹힘성은 대조구와 처리구간에 유의적인 차이를 보이지 않았다. 경도는 대조구와 젤라틴 1% 첨가구가 다른 처리구에 비해 유의적으로 낮았으며 (P<0.05), 젤라틴 5% 첨가구가 가장 높은 값을 나타내었다. 또한 검성은 젤라틴 5% 첨가구가 대조구에 비해 유의적으로 높은 값을 나타내었다(P<0.05). 이러한 결과로 보아 유화형 소시지에 젤라틴 첨가 시 이화학적 특성을 우수하게 변화시키는 것으로 나타내어 향후 다양한 소시지형 제품에 기능성 소재로 활용할 가능성이 있을 것으로 판단된다. The aim of this study was to investigate the effects of pork skin gelatin on the physicochemical properties (pH, CIE color value, water holding capacity, cooking yield, viscosity, proximate composition, and texture profile analysis) of emulsion-type sausage. Emulsion-type sausages were manufactured with 0% (control), 1%, 3%, and 5% pork skin gelatin. Moisture contents of samples containing 3% and 5% pork skin gelatin were significantly higher than those of the control and samples containing 1% pork skin gelatin (P<0.05). Protein contents were the highest in samples containing 5% pork skin gelatin (P<0.05). The pH values of uncooked and cooked samples increased with increasing pork skin gelatin level (P<0.05). The lightness and yellowness values of cooked samples containing pork skin gelatin were higher than those of the control (P<0.05). In addition, redness values of cooked samples containing 3% and 5% pork skin gelatin were significantly lower than those of the control and samples containing 1% pork skin gelatin (P<0.05). Water holding capacity (WHC) was the lowest in control, and samples containing 3% and 5% pork skin gelatin had significantly higher WHC compared to the other samples (P<0.05). Cooking yield of samples increased with increasing concentration of pork skin gelatin (P<0.05). Samples containing 3% and 5% pork skin gelatin showed higher viscosity than the control and sample containing 1% pork skin gelatin (P<0.05). Springiness, cohesiveness, and chewiness of samples were not significantly different among the samples. Hardness values of the control and sample containing 1% pork skin gelatin were lower than those of other samples (P<0.05), and samples containing 5% pork skin gelatin had the highest hardness (P<0.05). Gumminess of sample containing 5% pork skin gelatin was significantly higher than that of the control (P<0.05). The results show that pork skin gelatin could improve the potential of emulsion-type sausage physicochemical properties.

Interaction of Porcine Myofibrillar Proteins and Various Gelatins: Impacts on Gel Properties

Sin-Woo Noh,Dong-Heon Song,Youn-Kyung Ham,Tae-Kyung Kim,Yun-Sang Choi,Hyun-Wook Kim 한국축산식품학회 2019 한국축산식품학회지 Vol.39 No.2

The objectives of this study were to determine the interaction between porcine myofibrillar proteins and various gelatins (bovine hide, porcine skin, fish skin, and duck skin gelatins) and their impacts on gel properties of porcine myofibrillar proteins. Porcine myofibrillar protein was isolated from pork loin muscle (M. longissimus dorsi thoracis et lumborum). Control was prepared with only myofibrillar protein (60 mg/mL), and gelatin treatments were formulated with myofibrillar protein and each gelatin (9:1) at the same protein concentration. The myofibrillar protein-gelatin mixtures were heated from 10°C to 75°C (2°C/min). Little to no impacts of gelatin addition on pH value and color characteristics of heat-induced myofibrillar protein gels were observed (p>0.05). The addition of gelatin slightly decreased cooking yield of heat-induced myofibrillar protein gels, but the gels showed lower centrifugal weight loss compared to control (p<0.05). The addition of gelatin significantly decreased hardness, cohesiveness, gumminess, and chewiness of heat-induced myofibrillar gels. Further, sodium dodecyl poly-acrylamide gel electrophoresis (SDS-PAGE) showed no interaction between myofibrillar proteins and gelatin under non-thermal conditions. Only a slight change in the endothermic peak (probably myosin) of myofibrillar protein-gelatin mixtures was found. The results of this study show that the addition of gelatin attenuated the water-holding capacity and textural properties of heat-induced myofibrillar protein gel. Thus, it could be suggested that well-known positive impacts of gelatin on quality characteristics of processed meat products may be largely affected by the functional properties of gelatin per se, rather than its interaction with myofibrillar proteins.

아세트산 처리 황다랑어(Thunnus albacares ) 껍질 유래 젤라틴으로 제조한 바이오필름의 물리적 특성

김주연,김도형,김선봉 한국수산과학회 2011 한국수산과학회지 Vol.44 No.6

The present study examined the physical properties of biofilms manufactured from yellowfin tuna Thunnus albacares skin gelatin with the aim of developing a biofilm from fisheries by-products to replace mammalian sources. The physical properties of biofilms from yellowfin tuna gelatin were compared with those of biofilms from porcine gelatin. The yellowfin tuna gelatin biofilm exhibited higher tensile strength (69.08 MPa) and greater elongation (14.32%)than did porcine gelatin biofilm (50.50 MPa and 10.21%, respectively). The ΔE and YI (yellowness index) Huntercolor values of yellowfin tuna gelatin biofilm were three-fold and 15-fold higher, respectively, than values for porcine gelatin biofilm. The opacity value of yellowfin tuna gelatin biofilm was higher than that of porcine gelatin biofilm. The stability against water of yellowfin tuna gelatin biofilm was lower than that of porcine gelatin biofilm at pH 3 to pH 11. Thermogravimetric analysis (TGA) indicated that the thermal stability of the biofilms was about 270˚C for porcine gelatin biofilm and about 250˚C for yellowfin tuna gelatin biofilm.

제대혈에서 유핵세포 분리방법에 따른 회복률

이건수,김미정,최은진,최성민 대한조혈모세포이식학회 1998 대한조혈모세포이식학회지 Vol.3 No.2

연구배경 : 제대혈 조혈모세포 이식은 혈액 및 종양질환의 치료로 여러 가지의 이점으로 인해 많은 연구와 더불어 은행화에 대한 노력이 활발히 진행되고 있다. 그러나 이식의 성공률을 향상시키기 위한 제대혈 조혈모세포의 채집 방법과 획득, 장기간 냉동보존에 대해 표준화된 방법이 아직 제시되고 있지 못한 실정이다. 이 연구는 이러한 제대혈 조혈모세포의 획득률의 향상을 위해 시도되었다. 방법 : 건강한 산모로부터 제대혈 54례를 무균적, 노출(open) 방법으로 채혈하였다. F-H법, Gelatin법, HES법으로 적혈구 제거 또는 유핵세포를 분리하여 자동세포분석기를 이용하여 획득률을 구하였다. 세 가지 방법으로 분리된 유핵세포를 각각 2%, 10% 그리고 17% FBS 농도의 배지에서 실온에서 1시간, 2시간, 3시간, 20시간 그리고 24 시간 보관 후 회복률과 비교하였다. Gelatin법과 HES법으로 적혈구를 제거한 검체들은 세포수를 L-15배지로 2배 희석하여 각각 2%, 10% 그리고 17% FBS 농도의 배지에서 실온에서 1시간, 2시간, 3시간, 20시간 그리고 24 시간 보관 후 회복률을 확인하여 세포수를 2배 희석하기 전에 회복률과 비교하였다. 세 가지 세포분리법에 의해 얻어진 유핵세포를 프로그램냉동법을 이용하여 냉동한 후 37℃ 보육기에서 해동 후 회복률을 비교하였다. 결과 : 유핵세포분리에서 F-H법으로 분리한 총 백혈구의 획득률은 27.2%, Gelatin법 71.5%, HES법 73.7%로 Gelatin법이나 HES법이 F-H법에 비해 많았으나 다핵세포 제거율은 F-H법 92.9%, Gelatin법 38.2%, HES법 32.1%로 F-H법이 Gelatin법이나 HES법에 비해 통계적으로 유의하게 높았다.(P<0.05). 16례에서 실시된 3가지 방법에 의해 분리된 단핵세포 또는 유핵세포를 2% FBS가 섞인 L-15 배지에 희석한 후 보관시간에 따른 회복률의 비교에서 F-H법과 Gelatin법에 비해 HES법에 의한 세포분리방법에 의할 때 회복률은 시간경과에 따라 더 급격히 감소하였고, 10% FBS가 섞인 L-15 배지에 희석한 후 보관시간에 따른 회복률의 비교에서는 F-H법은 회복률이 유지되는데 비해 Gelatin법과 HES법에서는 회복률의 감소를 보이고 있으나 통계적인 유의성은 없었다. 17% FBS가 섞인 L-15 배지에 희석한 후 보관시간에 따른 회복률의 비교에서는 F-H법, Gelatin법 그리고 HES법에서 모두 회복률의 감소를 보였다. Gelatin법과 HES법에서 세포 수를 2배 희석하여 각각 2%, 10% 그리고 17%농도의 FBS가 섞인 L-15배지에 희석한 후 보관시간에 따른 회복률은 세포 수를 희석하기 전과 비교하여 통계적으로 유의한 차이를 보이지는 않았다. F-H법, Gelatin법, HES법으로 분리한 유핵세포를 프로그램냉동법으로 냉동하고 37℃에서 해동하였을 때 회복률은 각각 85.8%, 65.3%, 72.7%로 통계적으로 유의하기는 않았다. 결론 : 유핵세포분리는 Gelatin법이나 HES법이 F-H법에 비해 많았으나, 조혈모세포 이식에 불필요한 적혈구와 다핵세포의 제거율이 낮았고, 보관시간이 길수록 회복률의 감소가 현저했으며, F-H법에 의해 단핵세포를 분리한 검체를 10% FBS 농도의 배지에 보관했을 때 비교적 회복률이 유지되었다. 그러므로 세포분리 방법에 상관없이 냉동보존 전의 보관시간이 짧을수록 더 높은 회복률을 얻을 수 있었다. 프로그램냉동법을 이용하고 급속 해동을 실시할 경우 단핵세포 또는 유핵세포 분리방법에 따른 회복률의 차이는 없었다. Background: Human umbilical cord blood contains sufficient numbers of hematopoietic stem cells to provide long-term engraftment in the unrelated recipient with several advantages. The number of hematopoietic stem cells is an important factor for better results in transplantation. The purpose of this study is the development of a technique for the collection, separation and cryopreservation of the cord blood mononuclear cells for transplantation. Methods: Fifty four cord bloods were collected during deliveries at Kyungpook National University Hospital and Hana Hospital, Taegu, Korea from May, 1997 to May, 1998. The mononuclear cell separation by Ficoll-Hypaque (F-H) or red cell sedimentation by 3% Gelatin or 6% Hydroxyethyl Starch (HES) was done and the number and recovery rates were compared with hemocytometry. The recovery rates of separated cells by different cell separation methods in the L-15 medium containing 2%, 10% or 17% FBS before cryopreservation according to the suspension time were compared. The recovery rates of separated cells by Gelatin or HES which were diluted two times in the L-15 medium containing 2%, 10% or 17% FBS were compared as the same way. The recovery rates according to different cell separation methods after a programmed freezing method and thawing in the incubator were compared. Results: The mean volume of collected cord blood was 121 mL (80-155 mL). The removal of red blood cell was 99.4% by F-H, 79.6% by Gelatin and 70.3% by HES. The mononuclear cell separation was 52.6±27.2% by F-H, 91.1±28.9% by Gelatin and 89.4±27.8% by HES. The removal of polymorphonuclear cell was 92.9% by F-H, 38.2% by Gelatin and 32.1% by HES. The recovery rates of separated cells by Gelatin or HES in the L-15 medium containing 2%, 10% or 17% FBS before cryopreservation were gradually decreased according to the longer suspension time, and those by F-H in the L-15 medium containing 2% or 17% FBS before cryopreservation were also gradually decreased, but those by F-H in the L-15 medium containing 10% were relatively maintained up to 24 hours (P<0.01). The recovery rates of two times diluted separated cell concentration by Gelatin or HES in the L-15 medium containing 2%, 10% or 17% FBS were also decreased according to the longer suspension time as those of the initially separated cell concentration were done. The recovery rate after programmed freezing and thawing was 85.8% by F-H, 65.3% by Gelatin and 72.7% by HES. Conclusion: The nucleated cell separation by 3% Gelatin or 6% HES showed better yield than F-H. The red cell and polynuclear cell contaminations were higher in the former two methods. The recovery rates of separated cells were not significantly decreased in the 10% FBS by F-H method. The shorter the suspension time before cryopreservation, the better the recovery rate of separated cells regardless of cell separation methods. The programmed freezing showed good recovery rates of cells after thawing. These will be important factors in raising the success rate in hematopoietic stem cell transplantation.