摘要
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 (periostealosteoblast,POB)、颅骨成骨细胞 (cranialosteoblast,COB)和骨髓基质干细胞(marrowstromalcell,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 。
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.
出处
《中华创伤杂志》
CAS
CSCD
北大核心
2001年第1期10-13,共4页
Chinese Journal of Trauma
基金
国家重点基础研究发展规划 (973)项目<组织工程研究的科学问题>!(G19990 5 430 8)
国家自然科学基金重点资助项目!(39830 10 0 )
