CAJ | 학술논문

目的　比较从同一胚胎来源的骨膜成骨细胞、颅骨成骨细胞和骨髓基质干细胞的分裂增殖能力和细胞的功能状态。　方法　从人5个月龄胚胎骨膜、颅骨和骨髓组织中分离培养出骨膜成骨细胞（periosteal osteoblast, POB）、颅骨成骨细胞（cranial osteoblast, COB）和骨髓基质干细胞（marrow stromal cell, MSC），用第6代细胞进行比较研究。观察细胞形态和超微结构；通过细胞生长曲线和3H-胸腺嘧啶脱氧核苷(3H-TdR)掺入实验，比较细胞的分裂增殖能力；3H-脯氨酸(3H-Proline)掺入实验检测胶原合成能力；RT-PCR法检测Ⅰ型胶原、骨钙素mRNA表达等。并比较3种细胞的碱性磷酸酶(ALP)活性。　结果　3种细胞在形态上无显著区别。在超微结构上，POB和COB表现为高分化状态，MSC为低分化细胞；3种细胞的增殖能力依次为MSC>COB>POB；3种细胞的成骨能力(包括Ⅰ型胶原合成能力、骨钙素表达和ALP活性)依次为POB>COB>MSC。　结论　骨组织工程研究需要大量增殖能力强、功能旺盛、无限传代的种子细胞，3种来源的成骨细胞均不能满足骨组织工程研究要求，通过基因改造成骨细胞可能是一条有效途径。
목적　비교종동일배태래원적골막성골세포、로골성골세포화골수기질간세포적분렬증식능력화세포적공능상태。　방법　종인5개월령배태골막、로골화골수조직중분리배양출골막성골세포（periosteal osteoblast, POB）、로골성골세포（cranial osteoblast, COB）화골수기질간세포（marrow stromal cell, MSC），용제6대세포진행비교연구。관찰세포형태화초미결구；통과세포생장곡선화3H-흉선밀정탈양핵감(3H-TdR)참입실험，비교세포적분렬증식능력；3H-포안산(3H-Proline)참입실험검측효원합성능력；RT-PCR법검측Ⅰ형효원、골개소mRNA표체등。병비교3충세포적감성린산매(ALP)활성。　결과　3충세포재형태상무현저구별。재초미결구상，POB화COB표현위고분화상태，MSC위저분화세포；3충세포적증식능력의차위MSC>COB>POB；3충세포적성골능력(포괄Ⅰ형효원합성능력、골개소표체화ALP활성)의차위POB>COB>MSC。　결론　골조직공정연구수요대량증식능력강、공능왕성、무한전대적충자세포，3충래원적성골세포균불능만족골조직공정연구요구，통과기인개조성골세포가능시일조유효도경。
Objective　To investigate the proliferative and osteogenic activity of periosteal osteoblasts (POB), calvarial osteoblasts (COB) and bone marrow stromal cells (MSC) from the same embryo. Methods　POB, COB and MSC were isolated and cultured respectively in vitro from the periosteum, calvarium and bone marrow of 5-month embryos aborting spontaneously. The 6th-generation osteoblasts were studied comparatively. The morphological features of POB, COB and MSC were observed under microscope and transmission electron microscope. The proliferative abilities of POB, COB and MSC were detected respectively with growth curves and 3 H-TdR incorporation test, the collagen synthesis was studied with 3 H-proline incorporation test, and mRNA expressions of type Ⅰcollagen and osteocalcin were detected by RT-PCR technique, and the alkaline phosphatase (ALP) activities of POB, COB and MSC were studied comparatively. 　Results　Morphological characteristics of POB, COB and MSC had no obvious difference under microscope, and the organelles in POB were more well-developed than that of MSC and COB. From the results of growth curves and 3 H-TdR incorporation test, the proliferative ability of MSC was the strongest, and that of POB was the weakest. From the results of 3 H-proline incorporation test, mRNA expression of typeⅠcollagen and osteocalcin and ALP activity, the osteogenic activity of POB was the strongest, and that of MSC was the weakest.　Conclusions　POB, COB and MSC cannot meet the demand of bone tissue engineering. MSC has the strongest proliferative ability, but the osteogenic activity is too weak, while POB has strong osteogenic activity, but the proliferative ability is weak. Therefore, osteoblasts modified by gene transfection may be a better way out of the difficulty.