Hoechst 33258 Staining을 이용한 웅성 생쥐 성세포의 간편 분류

서지정보

ㆍ발행기관 : 한국수정란이식학회 ㆍ수록지정보 : Journal of Embryo Transfer / 30권 / 3호
ㆍ저자명 : 김경국, 박영식

목차

서 론
재료 및 방법
1. 공시동물
2. 주요 시약
3. 용액의 제조
4. 정원세포의 분리
5. 형광염색
6. 실험설계
7. 통계분석
결 과
1. 체외 발생 정자세포의 형광현미경 관찰
2. 체외발생 성세포 분류지표의 검정
고 찰
결 론
REFERENCES

영어 초록

In the study for a differentiation and development of spermatogonial cells, the researchers should commonly require a simple, fast and reasonable method that could evaluate the developmental stage of male germ cells without any damage and also relentlessly culture them so far as a cell stage aiming at experimental applications. For developing the efficient method to identify the stage of sperm cells, the morphological characteristics of sperm cells were investigated by staining the cells with blue fluorescent dye Hoechst 33258, and a criterion for male germ cell classification was elicited from results of the previous investigation, then the efficiency of the criterion was verified by applying it to assort the germ cells recovered from male mice in age from 6 to 35 days. As morphological characteristics, spermatogonia significantly differed from spermatocytes in size, appearance and fluorescent patches of nucleus, and spermatids could also be distinguished from spermatozoa by making a difference in the volume and shape of nucleus and the shape and fluorescence of tail. Aforesaid criterion was applicable for classifying in vitro cultured sperm cells by verifying its efficiency and propriety for assorting the stages of testicular germ cells. However, the fluorescent staining showed that germ cells in mouse testis should be dramatically differentiated and developed at 21 days and 35 days of age, which were known as times of sexual puberty and maturity in male mice, respectively. In conclusion, the results indicated that this simple criterion for sperm cell classification using fluorescence staining with Hoechst 33258 may be highly efficient and reasonable for spermatogenesis study.

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