1.Isolation of High Purity Mouse Mesenchymal Stem Cells through Depleting Macrophages Using Liposomal Clodronate
Ju Han SONG ; Jung-Woo KIM ; Mi Nam LEE ; Sin-Hye OH ; Xianyu PIAO ; Zhao WANG ; Seung-Hee KWON ; Ok-Su KIM ; Jeong-Tae KOH
Tissue Engineering and Regenerative Medicine 2022;19(3):565-575
BACKGROUND:
The use of mouse bone marrow mesenchymal stem cells (mBMSCs) represents a promising strategy for performing preclinical studies in the field of cell-based regenerative medicine; however, mBMSCs obtained via conventional isolation methods have two drawbacks, i.e., (i) they are heterogeneous due to frequent macrophage contamination, and (ii) they require long-term culturing for expansion.
METHODS:
In the present study, we report a novel strategy to generate highly pure mBMSCs using liposomal clodronate.This approach is based on the properties of the two cell populations, i.e., BMSCs (to adhere to the plasticware in culture dishes) and macrophages (to phagocytose liposomes).
RESULTS:
Liposomal clodronate added during the first passage of whole bone marrow culture was selectively engulfed by macrophages in the heterogeneous cell population, resulting in their effective elimination without affecting the MSCs.This method allowed the generation of numerous high-purity Sca-1 + CD44 + F4/80 - mBMSCs ([ 95%) with just one passaging. Comparative studies with mBMSCs obtained using conventional methods revealed that the mBMSCs obtained in the present study had remarkably improved experimental utilities, as demonstrated by in vitro multilineage differentiation and in vivo ectopic bone formation assays.
CONCLUSION
Our newly developed method, which enables the isolation of mBMSCs using simple and convenient protocol, will aid preclinical studies based on the use of MSCs.
2.Developmental Delay and Rehabilitation in an Infant with Partial Trisomy 1q32.1 to 1q44: A Case Report
Woo Kyung KIM ; Na Mi LEE ; In Seok LIM ; Soo Ahn CHAE ; Sin Weon YUN ; Dae Yong YI ; Su Yeong KIM
Neonatal Medicine 2022;29(2):91-95
Partial trisomy 1q is a rare chromosomal disorder characterized by ventriculomegaly with craniofacial, renal, cardiac, and finger and toe anomalies. Most reported cases of partial trisomy1q have involved stillborn or premature deaths due to cardiac or liver failure. This case report describes an 18-month-old patient with partial duplication of the 1q32-44 segments and consequent developmental delays who exhibited improvement in developmental status with rehabilitation. Prenatal ultrasonography and magnetic resonance imaging of the mother revealed ventriculomegaly and atrophic changes in the left cerebral hemisphere of the fetus. The infant was born with micrognathia, microphthalmia, macrocephaly, low-set ears, polydactyly, and long feet at 37+5 weeks of gestation. A chromosomal study revealed an abnormal male karyotype of 46,XY,rec(1)dup(1)(q32.1q44)inv(1)(p36.3q32.1)pat. In this rare case of a patient with partial trisomy, we observed improvement in developmental delays following treatment using appropriate rehabilitation techniques. Further research is required to help validate the findings of this case study and establish a standardized rehabilitation technique that can be subsequently applied to such cases.
3.Bone Generation Following Repeated Administration of Recombinant Bone Morphogenetic Protein 2
Hye-Ju SON ; Mi Nam LEE ; Yuri KIM ; Hyuck CHOI ; Byung-Chul JEONG ; Sin-Hye OH ; Jung-Woo KIM ; Seung-Hee KWON ; Sun-Hun KIM ; Soo-Chang SONG ; Shee Eun LEE ; Jeong-Tae KOH
Tissue Engineering and Regenerative Medicine 2021;18(1):155-164
BACKGROUND:
The delivery of recombinant human bone morphogenetic protein 2 (rhBMP2) by using various carriers has been used to successfully induce bone formation in many animal models. However, the effect of multiple administration of rhBMP2 on bone formation and BMP2 antibody production has not been determined. Our aim was to examine the bone formation activity of rhBMP2 and serum levels of anti-BMP2 antibodies following the repeated administration of rhBMP2 in mice.
METHODS:
Absorbable collagen sponges or polyphosphazene hydrogels containing rhBMP2 were subcutaneously implanted or injected into one side on the back of six-week-old C57BL/6 mice. Three or 4 weeks later, the same amount of rhBMP2 was administered again with the same carrier into the subcutaneous regions on the other side of the back or into calvarial defects. The effects of a single administration of rhBMP2 on the osteoinductive ability in the ectopic model were compared with those of repeated administrations. In vivo ectopic or orthotopic bone formation was evaluated using microradiography and histological analyses. Serum concentrations of anti-rhBMP2 antibodies were measured by ELISAs.
RESULTS:
Re-administration of the same amount of rhBMP2 into the subcutaneous area showed a comparable production of ectopic bone as after the first administration. The bone forming ability of repeated rhBMP2 administrations was equal to that of single rhBMP2 administration. The administration of rhBMP2 into calvarial defects, following the first subcutaneous administration of rhBMP2 on the back, completely recovered the defect area with newly regenerated bone within 3 weeks. Repeated administration of rhBMP2 at 4-week intervals did not significantly alter the serum levels of antiBMP2 antibodies and did not induce any inflammatory response. The serum obtained from rhBMP2-exposed mice had no effect on the ability of rhBMP2 to induce osteogenic gene expressions in MC3T3-E1.
CONCLUSION
We suggest that the osteoinductive ability of rhBMP2 is not compromised by repeated administrations. Thus, rhBMP2 can be repeatedly used for bone regeneration at various sites within a short duration.
4.Bone Generation Following Repeated Administration of Recombinant Bone Morphogenetic Protein 2
Hye-Ju SON ; Mi Nam LEE ; Yuri KIM ; Hyuck CHOI ; Byung-Chul JEONG ; Sin-Hye OH ; Jung-Woo KIM ; Seung-Hee KWON ; Sun-Hun KIM ; Soo-Chang SONG ; Shee Eun LEE ; Jeong-Tae KOH
Tissue Engineering and Regenerative Medicine 2021;18(1):155-164
BACKGROUND:
The delivery of recombinant human bone morphogenetic protein 2 (rhBMP2) by using various carriers has been used to successfully induce bone formation in many animal models. However, the effect of multiple administration of rhBMP2 on bone formation and BMP2 antibody production has not been determined. Our aim was to examine the bone formation activity of rhBMP2 and serum levels of anti-BMP2 antibodies following the repeated administration of rhBMP2 in mice.
METHODS:
Absorbable collagen sponges or polyphosphazene hydrogels containing rhBMP2 were subcutaneously implanted or injected into one side on the back of six-week-old C57BL/6 mice. Three or 4 weeks later, the same amount of rhBMP2 was administered again with the same carrier into the subcutaneous regions on the other side of the back or into calvarial defects. The effects of a single administration of rhBMP2 on the osteoinductive ability in the ectopic model were compared with those of repeated administrations. In vivo ectopic or orthotopic bone formation was evaluated using microradiography and histological analyses. Serum concentrations of anti-rhBMP2 antibodies were measured by ELISAs.
RESULTS:
Re-administration of the same amount of rhBMP2 into the subcutaneous area showed a comparable production of ectopic bone as after the first administration. The bone forming ability of repeated rhBMP2 administrations was equal to that of single rhBMP2 administration. The administration of rhBMP2 into calvarial defects, following the first subcutaneous administration of rhBMP2 on the back, completely recovered the defect area with newly regenerated bone within 3 weeks. Repeated administration of rhBMP2 at 4-week intervals did not significantly alter the serum levels of antiBMP2 antibodies and did not induce any inflammatory response. The serum obtained from rhBMP2-exposed mice had no effect on the ability of rhBMP2 to induce osteogenic gene expressions in MC3T3-E1.
CONCLUSION
We suggest that the osteoinductive ability of rhBMP2 is not compromised by repeated administrations. Thus, rhBMP2 can be repeatedly used for bone regeneration at various sites within a short duration.
5.CCR5 deficiency in aged mice causes a decrease in bone mass
Eun Ji OH ; Yaran ZANG ; Jung Woo KIM ; Mi Nam LEE ; Ju Han SONG ; Sin Hye OH ; Seung Hee KWON ; Jin Woo YANG ; Jeong Tae KOH
International Journal of Oral Biology 2019;44(4):173-181
The CC chemokine receptor 5 (CCR5) is a G protein-coupled receptor that regulates chemotaxis and effector functions of immune cells. It also serves as the major co-receptor for the entry of human immunodeficiency virus (HIV). Recently, CCR5 inhibitors have been developed and used for the treatment or prevention of HIV infections. Additionally, it has been identified that CCR5 controls bone homeostasis by regulating osteoclastogenesis and the communication between osteoblasts and osteoclasts. However, the effects of CCR5 inhibition on bone tissue in elderly patients are unknown. This study aimed to examine the bone phenotype of aged CCR5 knockout (KO) mice. Femoral and tibial bones were isolated from 12-month and 18-month old wild-type (WT) and CCR5 KO mice, and microcomputed tomography and histology analyses were performed. Twelve-month-old CCR5 KO mice exhibited a decreased trabecular bone mass and cortical bone thickness in both femoral and tibial bones compared with age-matched WT mice. Eighteen-month-old mice also showed a decreased trabecular bone mass in femurs compared with control WT mice, but not in tibial bones. Unlike in 12-month-old mice, the cortical margin of femurs and tibias in 18-month-old mice were rough, likely because they were aggravated by the deficiency of CCR5. Overall, our data suggest that the deficiency of CCR5 with aging can cause severe bone loss. When CCR5 inhibitors or CCR5 inactivating technologies are used in elderly patients, a preventive strategy for bone loss should be considered.
6.Identification of Somatic KRAS Mutation in a Korean Baby with Nevus Sebaceus Syndrome.
Sung Woo KIM ; Ju Sun SONG ; Mi Seon KANG ; Jong Beom SIN ; Chang Seok KI ; Ga Won JEON
Annals of Laboratory Medicine 2015;35(1):178-180
No abstract available.
Base Sequence
;
Child, Preschool
;
DNA/chemistry/metabolism
;
Female
;
Humans
;
Mutation
;
Nevus, Pigmented/diagnosis/*genetics
;
Polymorphism, Single Nucleotide
;
Proto-Oncogene Proteins p21(ras)/*genetics
;
Republic of Korea
;
Skin/pathology
;
Skin Neoplasms/diagnosis/*genetics
;
Syndrome
7.Neuroprotective effects of mild hypoxia in organotypic hippocampal slice cultures.
Seh Hyun KIM ; Woo Soon LEE ; Na Mi LEE ; Soo Ahn CHAE ; Sin Weon YUN
Korean Journal of Pediatrics 2015;58(4):142-147
PURPOSE: The aim of this study was to investigate the potential effects of mild hypoxia in the mature and immature brain. METHODS: We prepared organotypic slice cultures of the hippocampus and used hippocampal tissue cultures at 7 and 14 days in vitro (DIV) to represent the immature and mature brain, respectively. Tissue cultures were exposed to 10% oxygen for 60 minutes. Twenty-four hours after this hypoxic insult, propidium iodide fluorescence images were obtained, and the damaged areas in the cornu ammonis 1 (CA1), CA3, and dentate gyrus (DG) were measured using image analysis. RESULTS: In the 7-DIV group compared to control tissue, hypoxia-exposed tissue showed decreased damage in two regions (CA1: 5.59%+/-2.99% vs. 4.80%+/-1.37%, P=0.900; DG: 33.88%+/-12.53% vs. 15.98%+/-2.37%, P=0.166), but this decrease was not statistically significant. In the 14-DIV group, hypoxia-exposed tissue showed decreased damage compared to control tissues; this decrease was not significant in the CA3 (24.51%+/-6.05% vs. 18.31%+/-3.28%, P=0.373) or DG (15.72%+/-3.47% vs. 9.91%+/-2.11%, P=0.134), but was significant in the CA1 (50.91%+/-5.90% vs. 32.30%+/-3.34%, P=0.004). CONCLUSION: Although only CA1 tissues cultured for 14 DIV showed significantly less damage after exposure to hypoxia, the other tissues examined in this study showed a tendency towards less damage after hypoxic exposure. Therefore, mild hypoxia might play a protective role in the brain.
Anoxia*
;
Brain
;
Dentate Gyrus
;
Fluorescence
;
Hippocampus
;
Neuroprotective Agents*
;
Oxygen
;
Propidium
8.Validation of the Korean version of the European Community Respiratory Health Survey screening questionnaire for use in epidemiologic studies for adult asthma
Woo Jung SONG ; So Hee LEE ; Min Gyu KANG ; Ju Young KIM ; Mi Young KIM ; Eun Jung JO ; Suh Young LEE ; Seung Eun LEE ; Min Hye KIM ; Min Suk YANG ; Sae Hoon KIM ; Hye Ryun KANG ; Hye Kyung PARK ; Heung Woo PARK ; Yoon Seok CHANG ; Sun Sin KIM ; Jong Myung LEE ; Kyung Up MIN ; Sang Heon CHO
Asia Pacific Allergy 2015;5(1):25-31
BACKGROUND: Standardized questionnaire is one of key instruments for general population surveys. OBJECTIVE: The present study aimed to develop and validate the Korean version of the European Community Respiratory Health Survey (ECRHS) screening questionnaire for adult asthma surveys. METHODS: The ECRHS screening questionnaire was translated into Korean language according to the international criteria. Study participants were prospectively recruited from six referral hospitals and one health check-up center. Comprehensibility of the translation was tested in a pilot study of 10 patients. The reliability was evaluated by internal consistency and test-retest repeatability. Validity was assess with regard to physician-diagnosed asthma. RESULTS: A total of 100 adult asthma patients and 134 volunteers were recruited. Reliability was examined for 10 items in 100 asthmatics; Cronbach α coefficients were 0.84, and test-retest repeatability was good (Cohen κ coefficient, 0.71-1.00). Validity was assessed for 8 items in 234 participants; in particular, 'recent wheeze' showed a high sensitivity (0.89) for physician-diagnosed asthma. 'Recent asthma attack' and 'current asthma medication' showed high specificity (0.96-0.98). CONCLUSION: The present study demonstrated that the Korean version of the ECRHS screening questionnaire was comprehensible, reliable and valid. We suggest the questionnaire to be utilized in further epidemiological studies for asthma in Korean adult populations.
Adult
;
Asthma
;
Epidemiologic Studies
;
Epidemiology
;
European Union
;
Health Surveys
;
Humans
;
Mass Screening
;
Pilot Projects
;
Prospective Studies
;
Referral and Consultation
;
Sensitivity and Specificity
;
Volunteers
9.Changes in Hypermetropic Spectacle Correction after Surgery in Partially Accommodative Esotropia.
Sin Woo BAE ; Moses KIM ; Mi Young CHOI
Journal of the Korean Ophthalmological Society 2014;55(5):719-725
PURPOSE: To investigate the characteristics of successfully weaning patients off of glasses and the change in hypermetropic spectacle correction required for maintaining orthotropia using an analysis of surgery results of patients with partially accommodative esotropia. METHODS: We reviewed the medical records of 104 patients who underwent standard surgery for correcting partially accommodative esotropia. In total, 64 patients who had follow-up periods of at least 2 years were included. The patients were divided into 2 groups: 28 patients who were asked to discontinue their hyperopic glasses (glasses-discontinued group) and 36 patients who still needed hyperopic glasses (glasses-maintained group). We investigated the age at first visit and at surgery, total angle of deviation and residual angle of deviation with correction before surgery, weaning time of hyperopic glasses, follow-up period, and the time at which the hyperopic glasses were discontinued in the glasses-discontinued group. RESULTS: There were no statistically significant differences in the age at first visit and at surgery as well as the duration of postoperative follow-up between both groups. The total esodeviated angle without hyperopic correction of the glasses-discontinued group was significantly lower than that of glasses-maintained group (37.4PD:46.7PD, p < 0.05); there were no significant differences in the remaining esotropic angle after hyperopic correction. The average hyperopic degree in the glasses-discontinued group was significantly lower than that in the glasses-maintained group (+3.0D:+4.7D, p < 0.05), there were no significant differences in the weaning time of hyperopic glasses between both groups. CONCLUSIONS: The esodeviated angle without hyperopic correction was smaller and hyperopic degree was lower in the glasses-discontinued group than in the glasses-maintained group after surgery for partially accommodative esotropia. Therefore, it might be helpful to predict the postoperative possibility to discontinue glasses in the patients with partially accommodative esotropia.
Esotropia*
;
Eyeglasses
;
Follow-Up Studies
;
Glass
;
Humans
;
Hyperopia
;
Medical Records
;
Weaning
10.Changes in Hypermetropic Spectacle Correction after Surgery in Partially Accommodative Esotropia.
Sin Woo BAE ; Moses KIM ; Mi Young CHOI
Journal of the Korean Ophthalmological Society 2014;55(5):719-725
PURPOSE: To investigate the characteristics of successfully weaning patients off of glasses and the change in hypermetropic spectacle correction required for maintaining orthotropia using an analysis of surgery results of patients with partially accommodative esotropia. METHODS: We reviewed the medical records of 104 patients who underwent standard surgery for correcting partially accommodative esotropia. In total, 64 patients who had follow-up periods of at least 2 years were included. The patients were divided into 2 groups: 28 patients who were asked to discontinue their hyperopic glasses (glasses-discontinued group) and 36 patients who still needed hyperopic glasses (glasses-maintained group). We investigated the age at first visit and at surgery, total angle of deviation and residual angle of deviation with correction before surgery, weaning time of hyperopic glasses, follow-up period, and the time at which the hyperopic glasses were discontinued in the glasses-discontinued group. RESULTS: There were no statistically significant differences in the age at first visit and at surgery as well as the duration of postoperative follow-up between both groups. The total esodeviated angle without hyperopic correction of the glasses-discontinued group was significantly lower than that of glasses-maintained group (37.4PD:46.7PD, p < 0.05); there were no significant differences in the remaining esotropic angle after hyperopic correction. The average hyperopic degree in the glasses-discontinued group was significantly lower than that in the glasses-maintained group (+3.0D:+4.7D, p < 0.05), there were no significant differences in the weaning time of hyperopic glasses between both groups. CONCLUSIONS: The esodeviated angle without hyperopic correction was smaller and hyperopic degree was lower in the glasses-discontinued group than in the glasses-maintained group after surgery for partially accommodative esotropia. Therefore, it might be helpful to predict the postoperative possibility to discontinue glasses in the patients with partially accommodative esotropia.
Esotropia*
;
Eyeglasses
;
Follow-Up Studies
;
Glass
;
Humans
;
Hyperopia
;
Medical Records
;
Weaning

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