This study was carried out to investigate effective condition for producing somatic cell nuclear transfer (SCNT) embryos of Jeju native cattle. As donor cells for SCNT, ear skin cells from Jeju native cattle were used. In experiment 1, the effect of recipient oocyte sources on the development of Jeju native cattle SCNT embryos were examined. Fusion rate of recipient oocyte and donor cell was not different between the Hanwoo and Holstein recipient oocytes (86.0% vs 89.9%). The rate of embryos developing to the blastocyst stage was significantly (p<0.05) higher in Hanwoo recipient oocytes than in Holstein recipient ones (28.2% vs 14.7%). Blastocysts derived from Hanwoo recipient oocytes contained higher numbers of total cells than those derived from Holstein ones ($115.1{\pm}40.8$ vs $101.4{\pm}33.3$), although there were no significant difference. The mean proportion of apoptotic cells in blastocyst was not different between the sources of recipient oocytes. In experiment 2, the development of Jeju native cattle and Hanwoo SCNT embryos were compared. Hanwoo oocytes were used as the recipient oocytes. Fusion rate was not different between the Jeju native cattle and Hanwoo SCNT embryos (92.1% vs 92.9%). The blastocyst rate of SCNT embryos was significantly (p<0.05) lower in Jeju native cattle than in Hanwoo (16.9% vs 31.0%). Blastocysts derived from Jeju native cattle SCNT embryos contained smaller numbers of total cells than those derived from Hanwoo ones ($136.6{\pm}33.7$ vs $149.9{\pm}39.7$), but there were no significant difference. The mean proportion of apoptotic cells in blastocyst was not different between the Jeju native cattle and Hanwoo SCNT embryos. The present study demonstrated that Hanwoo recipient oocytes were more effective in supporting production of Jeju native cattle SCNT embryos, although Jeju native cattle SCNT embryos showed reduced developmental capacity when compared to Hanwoo SCNT embryos.
To improve the efficiency of production of cloned embryos and animals by nuclear transplantation in the rabbit, the effect of cell cycle of donor nuclei and type of recipient cytoplasm on the in vitro developmental potential and production efficiency of offspring was determined. The embryos of 16-cell stage were collected from the mated does at 48h post-hCG injection and they were synchronized to G$_1$ phase of 32-cell stage. The oocytes collected at 14h post-hCG injection were freed from cumulus cells and then enucleated. One group of the enucleated cytoplasms was activated by electrical stimulation prior to injection of donor nucleus, and the other group was not pre-activated. The separated G$_1$phase blastomeres of 32-cell stage embryos were injected into the perivitelline space of recipient cytoplasms. After culture for 20h post-hCG injection, the nuclear transplant oocytes were electrofused and activated by electrical stimulation and the fused nuclear transplant embryos were co-cultured for 120h and the nuclear transplant embryos developed to blastocyst stage were stained with Hoechst 33342 dye and their blastomeres were counted. Some of the nuclear transplant embryos developed in vitro to 2- to 4-cell stage were transferred into the oviducts of synchronized recipient does. The electrofusion rate was similar between the types of donor nuclei and recipient cytoplasms used. However, the nuclear transplant embryos using G$_1$ phase donor nuclei were developed to blastocyst at higher rate(60.3%) than those using S phase ones(24.7%). Also, when non-preactivated oocytes were used as recipient cytplasms, the develop-mental rates of nuclear transplant embryos to blastocysts were significantly(P< 0.05) higher(57.1%) than those using preactivated ones(20.8%). The cell counts of nuclear transplant embryos developed to blastosyst stage were increased signficantly(P<0.05) more in the non-preactivated recipient cytoplasm(163.7 cells), as compared whit the preactivated recipient cytoplasm(85.4 cells), A total of 49 nuclear transplant embryos were tranferrid into 5 recipient does, of which two offsprings were produced from a foster mother 31 days after embryo transfer. these results showed that the blastomeres of G1 phase and non-preactivated oocytes might be utillzed efficiently as donor nuclei and recipient cytoplasms in the nuclear transplant procedure, thought the offspring production remained still low.
Ahn, Hee Chang;Lee, Han Earl;Kim, Jeong Tae;Choi, M.Seung Suk
Archives of Plastic Surgery
/
v.34
no.5
/
pp.569-573
/
2007
Purpose: The selection of the recipient vessels in breast reconstruction has a great influence on the surgical result and the shape of the reconstructed breast. We would like to introduce the criteria for the selection of recipient vessels in delayed reconstruction of the breast. Methods: We studied 56 patients with delayed breast reconstruction using free TRAM flaps from April 1994 to December 2006. The thoracodorsal and the ipsilateral internal mammary vessels were used as recipients in 25 patients each, the opposite internal mammary vessels in 3 patients, the thoracoacromial vessels in 2 patients, and the transverse cervical artery with the cephalic vein in 1 patient. The survival rate of the flaps, the vessel diameter, the length of the pedicles, and the convenience of vessel dissection were studied. Results: The diameter of the recipient vessel did not influence the anastomosis. The operation time, the survival rate of flap, the postoperative complications showed no significant difference according to the recipient vessel. Dissection of the thoracodorsal vessels was tedious due to scar formation from the prior operation. Dissection of the internal mammary vessels proved to be relatively easy, and the required length of the pedicle was shorter than any other site, but the need for removal of rib cartilage makes this procedure inconvenient. Conclusion: The first choice of the recipient vessel in immediate breast reconstruction is the thoracodorsal vessels, but in cases of delayed reconstruction the internal mammary vessels are favored as the first choice, because the thoracodorsal vessels have a high unusability rate. If the ipsilateral internal mammary vessels prove to be useless, the contralateral vessels can be used. The thoracoacromial vessels are useful, when the mastectomy scar is located in the upper portion. The transverse cervical artery and the cephalic vein can serve as the last resort, if all other vessels are unreliable.
Park, Myong-Chul;Lee, Jung-Hoon;Chung, Jae-Ho;Lee, Sung-Hun
Archives of Reconstructive Microsurgery
/
v.10
no.2
/
pp.105-110
/
2001
Breast reconstruction is an aesthetically critical procedure and should be peformed to match the opposite breast in shape, contour, and position. Many methods were introduced to reconstruct the breast with autogenous tissue. But, free tissue transfer for breast reconstruction has become common method. The transverse rectus abdominis myocutaneous flap technique has been a widely accepted method of breast reconstruction after mastectomy, since the first introduction of free abdominoplasty flap in 1979. In breast reconstruction with a free flap the selection of suitable recipient vessels remains one of the most critical decision for surgeon. The most common recipient site for free flap breast reconstruction is the axillar system. But, the use of the axillary system as a recipient site limits flap movement and flexibility in breast shaping. The use of internal mammary vessels as a recipient site be able to achieve ideal breast symmetry, but that technique require the rib resection. The selection of suitable recipient vessels is most important for successful free tissue transfer. We have performed breast reconstruction with TRAM flaps anastomozed to the internal mammary vessel perforator. We came to the conclusion that this vessel perforator is useful as a recipient site in cases of immediate breast reconstruction with free TRAM flap.
A powerful tool for chicken transgenesis could be established by employing a germline chimera production through primordial germ cell transplantation. This study was conducted to examine whether foreign gene-transfected gonadal primordial germ cells (gPGCs) have a migration activity into the gonad after transfer to recipient embryos. In Experiment 1, gPGCs of Korean Ogol Chicken were retrieved from 5.5-day-old embryos and subsequently transferred to the dorsal aorta of 2.5-day-old White Leghorn embryos after being labeled with PKH26 fluorescent dye. To confirm migration activity after transplantation, recipient embryos were sacrificed and examined on 3 days after transfer. Sex determination was concomitantly undertaken to examine whether sex of recipient embryos could affect the migration activity of gPGCs. All of embryonic gonads examined showed positive signals with PKH26 fluorescence and W-chromosome specific band by polymerase chain reaction (PCR) was detected in male embryos when gPGCs with ZW chromosome were transferred to recipient embryos. In Experiment 2, retrieved gPGCs were transfected with LacZ gene-containing cytomegalovirus promoter ($pCMV{\beta}$) by electroporation and subsequently transferred to recipient embryos. LacZ gene expression was identified in the gonads of 6 or 10-day-old recipient embryos and hatched-chicks. A total of 20 embryos and 12 hatched-chicks were examined and 11 of them (10 embryos and one hatched chicken; 11/32=34.4%) expressed $\beta$-galactosidase, a marker substance of LacZ gene. The results of this study demonstrated that foreign gene-transfected gPGCs can migrate and settle down into the gonad after being transferred into the blood vessel of the recipient embryos. This established technique will contribute to developing a peer biotechnology for transgenic chicken.
Kim, Dong-Hoon;Kim, Se-Woong;Lee, Min-Jung;Bae, Seong-Hoon;Im, Gi-Sun;Lim, Hyun-Joo;Yang, Byoung-Chul;Seong, Hwan-Hoo
Reproductive and Developmental Biology
/
v.32
no.3
/
pp.175-181
/
2008
This study was conducted to investigate an effective recipient oocyte and culture system for producing of Hanwoo (Korean native cattle) somatic cell nuclear transfer (SCNT) embryos. Hanwoo ear skin fibroblasts were used as donor cells. In vitro matured Hanwoo or Holstein oocytes were enucleated, and single donor cells were transferred into the perivitelline space of the enucleated oocytes. The couplets were subsequently fused and activated. The reconstructed embryos were cultured in a conventional or sequential culture system. In the former, embryos were cultured in CR2aa medium for eight days; in the latter, embryos were cultured in modified CR2aa-A (mCR2-A) for three days and then further cultured in modified CR2aa-B (mCR2-B) for five days. In the experiment with the recipient oocyte, the rate of embryo development to the blastocyst stage was significantly (p<0.05) higher in Hanwoo recipient oocytes than in Holstein ones (48.8% vs 38.9%). BIastocysts derived from Hanwoo recipient oocytes contained significantly (p<0.05) higher numbers of total cells than those derived from Holstein recipient oocytes ($156.0{\pm}68.2$ vs $134.7{\pm}54.8$). There was no difference in the mean proportion of apoptotic cells in blastocysts between the sources of recipient oocytes. In the experiment with the embryo culture system, the blastocyst rate was somewhat higher in sequential system than in conventional system (50.0% vs 43.5%), though there was no significant difference. The numbers of total ($160.0{\pm}69.0$ vs $156.7{\pm}68.4$) and apoptotic cells ($14.0{\pm}10.4$ vs $11.8{\pm}6.4$) were not different between the culture systems. In conclusion, the present study demonstrated that Hanwoo recipient oocytes and the sequential culture system were more effective in supporting the production of Hanwoo SCNT embryos.
Kim, Byung-Gak;Lee, Yong-An;Kim, Bang-Jin;Kim, Ki-Jung;Min, Kwan-Sik;Lee, Jang-Hee;Ryu, Jae-Weon;Kim, In-Cheul;Ryu, Buom-Yong
Reproductive and Developmental Biology
/
v.32
no.3
/
pp.193-198
/
2008
The current study was designed to extend the technique of spermatogonial transplantation to economically important pig model We evaluated the efficiency of pig to pig transplantation. Donor testis cells were harvested from testes obtained at castration of 10- to 14-day-old boars and were labeled with fluorescent marker(PKH26) before transplantation. The presence of infused dye or labeled pig testicular cells was confirmed in the seminiferous tubules from recipient pig. The most effective procedure of intratubular germ cell transfer was to insert an fine needle ($21{\sim}25$ gauge) through the cauda epididymis and testis into the rete testis under ultrasound guidance. Infusion of $5{\sim}7ml$ of dye solution or cell suspension could fill the rete and up to 50% of seminiferous tubules of 14-week-old boars. Testis were examined for the presence and localization of labeled donor cells immediately after transplantation and labeled donor cells were found in numerous seminiferous tubules from recipient pig testes. These results indicate that germ cell transplantation is feasible in recipient pig testis. This study represents successful spermatogonial transplantation between individual animals in a livestock species.
Most research on Official Development Assistance (ODA) targeted the recipients, but this study examines the effect of ODA on the donor's exports to the recipient. To do this, a panel analysis was carried out with ODA and macroeconomic data on the United States and 33 other countries from 1999 to 2009. The results are summarized as follows: (1) The economic influence on the donor varies with the recipient's localization effort. (2) High-tech exports to the recipient are independent of ODA. (3) In medium-to-low-technology areas, ODA has a positive effect on exports to the recipient with low absorption efforts. (4) Both High-tech and LM-tech product exports decreases with the technological innovation efforts of recipients with high absorption efforts, while High-tech product exports only increases with that of recipients with low absorption efforts. These results indicate that a strategic approach for ODA program is more effective and useful to the donor's economy.
Multiplex PCR-based short tandem repeat (STR) analysis is considered as a good tool for monitoring bone marrow engraftment after sex-mismatched allogeneic transplantation and provides a sensitive and accurate assessment of the contribution of both donor and/or recipient cells in post-transplantation specimens. Forensic STR analysis and quantitative real time PCR are used to determine the proportion of donor versus recipient each contained within the total DNA. The STR markers were co-amplified in a single reaction by using commercial $PowerPlex^{(R)}$ 16 system and $AmpFISTR^{(R)}$$Identifiler^{(R)}$ / $Yfiler^{(R)}$ PCR amplification kits. Separation of the PCR products and fluorescence detection were performed by ABI $PRIS^{(R)}$ 3100 Genetic Analyzer with capillary electrophoresis. The $GeneMapper^{TM}$ ID software were used for size calling and analysis of STR profiles. Extracted DNA was quantified by the $Quantifiler^{TM}$ Human DNA / Y Human Male DNA Quantification Kit The intent of this study was to analyze the ratio of donor versus recipient cells in the post-transplant peripheral blood, spleen, lung and kidney specimens. Specimens were taken from the traffic accident male victim who had been engrafted from bone marrow female donor. Blood and spleen specimens displayed female donor DNA profile. Kidney specimen showed male recipient DNA profile. Interestingly, lung tissue showed mixed profiles. The findings of this study indicate that the forensic STR analysis using fluorescence labeling PCR combined with capillary electrophoresis is quick and reliable enough to assess the ratio of donor versus recipient cells and to monitor the mixed chimeric patterns.
In lower extremity reconstruction, the recipient vessel often requires long-range mechanical dilation because of extensive vasospasm or plaque formation induced by concomitant atherosclerosis. While a forceps dilator can be used to manipulate and dilate vessels approximately 1 cm from their end, a DeBakey vascular dilator can dilate long-range vessels. The authors successfully performed free flap reconstruction of the lower extremity using the DeBakey vascular dilator. Of the two patients who underwent lower extremity reconstruction, one had extensive vasospasm, and the other had plaques in the recipient arteries. Irrigation with 4% lidocaine and dilation of the lumen with a forceps dilator were insufficient to restore the normal arterial blood flow. Instead, a DeBakey vascular dilator with a 1-mm diameter tip was gently inserted into the lumen. Then, to overcome vessel resistance, the dilator gently advanced approximately 10 cm to dilate the recipient artery. Normal arterial blood flow was gushed out after dilating the vessel lumen using a DeBakey vascular dilator. The vascular anastomosis was performed, and intravenous heparin 5000 IU was administered immediately after anastomosis. Prophylactic low-molecular-weight-heparin (Clexane, 1 mg/kg) was administered subcutaneously to both patients for 14 days. The reconstructed flap survived without necrosis in either patient.
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